6.3.1.19: prokaryotic ubiquitin-like protein ligase
This is an abbreviated version!
For detailed information about prokaryotic ubiquitin-like protein ligase, go to the full flat file.
Word Map on EC 6.3.1.19
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6.3.1.19
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mycobacterium
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tuberculosis
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depupylation
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pup-proteasome
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actinobacteria
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depupylase
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deamidation
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ligases
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deamidase
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smegmatis
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ubiquitin-proteasome
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tcmsp
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cytoscape
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isopeptide
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genecards
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nitrospirae
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drug development
- 6.3.1.19
- mycobacterium
- tuberculosis
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depupylation
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pup-proteasome
- actinobacteria
- depupylase
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deamidation
- ligases
- deamidase
- smegmatis
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ubiquitin-proteasome
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tcmsp
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cytoscape
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isopeptide
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genecards
- nitrospirae
- drug development
Reaction
Synonyms
PafA, proteasome accessory factor A, Pup ligase, Pup-protein ligase, Rv2097c, ubiquitin-like protein ligase
ECTree
6.3.1.19 prokaryotic ubiquitin-like protein ligaseAdvanced search results
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results (Substrates Products
Substrates Products on EC 6.3.1.19 - prokaryotic ubiquitin-like protein ligase
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SUBSTRATE 
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [adenylate kinase-His6]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[adenylate kinase-His6]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [FabD]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[FabD]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [IdeR]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[IdeR]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [inositol 1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[inositol 1-phosphate synthetase]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [Log]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[Log]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [malonyl Co-A acyl carrier protein transacylase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[malonyl Co-A acyl carrier protein transacylase]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [myo-inositol-1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[myo-inositol-1-phosphate synthetase]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [prokaryotic ubiquitin-like protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[prokaryotic ubiquitin-like protein]-L-lysine
in addition to the lysine at position 61, Pup (prokaryotic ubiquitin-like protein) presents two more lysines, one at position 7 and another at position 31. Pup can be pupylated on different lysines
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [proteasome-interacting ATPase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[proteasome-interacting ATPase]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-[[prokaryotic ubiquitin-like protein]-L-glutamate]-[[protein]-L-lysine]
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [adenylate kinase-His6]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[adenylate kinase-His6]-L-lysine
adenylate kinase from Escherichia coli is pupylated
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [adenylate kinase-His6]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[adenylate kinase-His6]-L-lysine
adenylate kinase from Escherichia coli is pupylated
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [FabD]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[FabD]-L-lysine
FabD from Mycobacterium tuberculosis is pupylated on a preferred Lys173
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [FabD]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[FabD]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
FabD from Mycobacterium tuberculosis is pupylated on a preferred Lys173
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [IdeR]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[IdeR]-L-lysine
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [IdeR]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[IdeR]-L-lysine
Mycobacterium smegmatis recombinant His6-tagged IdeR is a model substrate for in vitro studies
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [IdeR]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[IdeR]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
Mycobacterium smegmatis recombinant His6-tagged IdeR is a model substrate for in vitro studies
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [IdeR]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[IdeR]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [inositol 1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[inositol 1-phosphate synthetase]-L-lysine
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [inositol 1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[inositol 1-phosphate synthetase]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
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-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [Log]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[Log]-L-lysine
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-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [Log]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[Log]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [malonyl Co-A acyl carrier protein transacylase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[malonyl Co-A acyl carrier protein transacylase]-L-lysine
Lys173 is the predominant modified residue in malonyl Co-A acyl carrier protein transacylase (FabD) purified from Escherichia coli. Lys122 and Lys181 are also identified as pupylation targets. At least three lysines in Mtb FabD can be pupylated in Escherichia coli. It also seems that only one residue per FabD polypeptide is pupylated at one time
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [malonyl Co-A acyl carrier protein transacylase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[malonyl Co-A acyl carrier protein transacylase]-L-lysine
Lys173 is the predominant modified residue in malonyl Co-A acyl carrier protein transacylase (FabD) purified from Escherichia coli. Lys122 and Lys181 are also identified as pupylation targets. At least three lysines in Mtb FabD can be pupylated in Escherichia coli. It also seems that only one residue per FabD polypeptide is pupylated at one time
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [myo-inositol-1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[myo-inositol-1-phosphate synthetase]-L-lysine
expression of Mycobacterium tuberculosis ino1, pafA and pupGlu, results in pupylated myo-inositol-1-phosphate synthetase (Ino1) in Escherichia coli
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [myo-inositol-1-phosphate synthetase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[myo-inositol-1-phosphate synthetase]-L-lysine
expression of Mycobacterium tuberculosis ino1, pafA and pupGlu, results in pupylated myo-inositol-1-phosphate synthetase (Ino1) in Escherichia coli
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
Mycobacterium tuberculosis PanB is a model substrate of PafA, in silico docking analysis reveals interaction via PafA residue arginine 207
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
Mycobacterium tuberculosis PanB is a model substrate of PafA, in silico docking analysis reveals interaction via PafA residue arginine 207
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
PanB from Mycobacterium tuberculosis is pupylated at a single lysine residue, K212, formation of a covalent MtbPup-MtbPanB conjugate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
PanB from Mycobacterium tuberculosis is pupylated at a single lysine residue, K212, formation of a covalent MtbPup-MtbPanB conjugate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
PanB is a model substrate of PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[PanB]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[phosphoenolpyruvate-protein phosphotransferase I]-L-lysine
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-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [proteasome-interacting ATPase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[proteasome-interacting ATPase]-L-lysine
pupylation of proteasome-interacting ATPase (Mpa) occurs predominantly on one target lysine. Pupylation at this position prevents interaction of Mpa/prokaryotic ubiquitin-like protein(Pup) with the proteasome core. Ultimately, pupylation leads to deoligomerization of the Mpa hexamer driven by the unfolding activity of Mpa, thereby rendering Mpa-Pup fully inactive
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [proteasome-interacting ATPase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[proteasome-interacting ATPase]-L-lysine
pupylation of proteasome-interacting ATPase (Mpa) occurs predominantly at Lys591
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [proteasome-interacting ATPase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[proteasome-interacting ATPase]-L-lysine
pupylation of proteasome-interacting ATPase (Mpa) occurs predominantly on one target lysine. Pupylation at this position prevents interaction of Mpa/prokaryotic ubiquitin-like protein(Pup) with the proteasome core. Ultimately, pupylation leads to deoligomerization of the Mpa hexamer driven by the unfolding activity of Mpa, thereby rendering Mpa-Pup fully inactive
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [proteasome-interacting ATPase]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[proteasome-interacting ATPase]-L-lysine
pupylation of proteasome-interacting ATPase (Mpa) occurs predominantly at Lys591
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
a pupylation product with a molecular mass approximately 7 kDa heavier than PanB suggests that each PanB monomer is singly pupylated by PafA. PanB is pupylated primarily on either one of two lysines, namely Lys34 or Lys36. Pupylation on Lys137 is also detected, although less frequently
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
the C-terminal glutamate of Pup-GGE is coupled to the protein substrate PanB lysine residue via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
[protein PanB]-L-lysine shows a 3 orders of magnitude stronger affinity than free lysine
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
the C-terminal glutamate of Pup-GGE is coupled to the protein substrate PanB lysine residue via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
[protein PanB]-L-lysine shows a 3 orders of magnitude stronger affinity than free lysine
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein PanB]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein PanB]-L-lysine
a pupylation product with a molecular mass approximately 7 kDa heavier than PanB suggests that each PanB monomer is singly pupylated by PafA. PanB is pupylated primarily on either one of two lysines, namely Lys34 or Lys36. Pupylation on Lys137 is also detected, although less frequently
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
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-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Acidothermus cellulolyticus ATCC 43068 / 11B
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Acidothermus cellulolyticus ATCC 43068 / 11B
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Bifidobacterium adolescentis ATCC 15703 / DSM 20083 / NCTC 11814 / E194a
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Bifidobacterium adolescentis ATCC 15703 / DSM 20083 / NCTC 11814 / E194a
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Mycobacterium smegmatis Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
the Mycobacterium smegmatis Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Kocuria rhizophila ATCC 9341 / DSM 348 / NBRC 103217 / DC2201
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Kocuria rhizophila ATCC 9341 / DSM 348 / NBRC 103217 / DC2201
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Micrococcus luteus ATCC 4698 / DSM 20030 / JCM 1464 / NBRC 3333 / NCIMB 9278 / NCTC 2665 / VKM Ac-2230
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Micrococcus luteus ATCC 4698 / DSM 20030 / JCM 1464 / NBRC 3333 / NCIMB 9278 / NCTC 2665 / VKM Ac-2230
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
key enzyme in the Pup tagging (i.e. pupylation) system. Protein pupylation can be regulated at the enzyme level via an allosteric mechanism
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
pupylation is a signal for proteasomal degradation in bacteria. The prokaryotic, ubiquitin-like protein (Pup) is conjugated through its C-terminal residue to lysine side chains of substrates via an isopeptide bond. Pup proteins are small, ranging from 60 to 70 residues in length with Gly-Gly-Glu or Gly-Gly-Gln as C-terminal residues. Deamidation of the C-terminal glutamine of Pup by the deamidase of Pup (Dop) renders Pup competent for conjugation to substrates, which is then carried out by the Pup-ligase PafA
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
no activity with ATPgammaS
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
PafA is an allosteric enzyme that binds its target substrates cooperatively. In contrast, binding of deamidated Pup (Pup-L-glutamate) by PafA follows simple kinetics and is, therefore, not involved in the allosteric transitions of PafA. Pup and target substrate binding by PafA are independent events
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Pup is linked to substrate proteins by the formation of an isopeptide bond between the side-chain carboxylate of glutamate64 and a lysine side chain of the substrate protein
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the C-terminal 26 amino acid sequence of the prokaryotic ubiquitin-like protein (Pup) is the minimal ligase recognition motif in Mycobacterium tuberculosis. Specific hydrophobic residues within this sequence that are known to be important for the interactions of Pup with proteasomes are also critical for the activation of Pup by PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the enzyme catalyzes a two-step reaction mechanism proceeding through a gamma-glutamyl phosphate-mixed anhydride intermediate that is formed on the C-terminal glutamate of the prokaryotic ubiquitin-like protein (Pup) before transfer of Pup to the substrate acceptor lysine. Phosphorylated [prokaryotic ubiquitin-like protein]-L-glutamate and ADP are retained on the enzyme in a stable fashion following activation
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the prokaryotic, ubiquitin-like protein (Pup) is conjugated through its C-terminal residue to lysine side chains of substrates via an isopeptide bond. Pup proteins are small, ranging from 60 to 70 residues in length with Gly-Gly-Glu or Gly-Gly-Gln as C-terminal residues. Deamidation of the C-terminal glutamine of Pup by the deamidase of Pup (Dop) renders Pup competent for conjugation to substrates, which is then carried out by the Pup-ligase PafA. The enzyme turns over one molecule of ATP to ADP per conjugation cycle, suggesting that PafA activates Pup by phosphorylation of its C-terminal glutamate. The formation of this intermediate occurs even in the absence of substrate and is shown to be the rate-limiting step of the PafA catalyzed reaction. After activation, the phosphorylated Pup and ADP remain bound to PafA, awaiting the nucleophilic attack of the substrate lysine, which finally results in the formation of the isopeptide bond. The C-terminal residue of the ligation-competent PupE features two carboxylates, the C-terminal alpha-carboxylate and the gamma-carboxylate of the glutamyl side chain, both of which could in principle be used for Pup-attachment. NMR-experiments using the pupylated proteasomal substrate PanB show that only the gamma-carboxylate is used to form the isopeptide bond between Pup and the target
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
pupylation is a signal for proteasomal degradation in bacteria. The prokaryotic, ubiquitin-like protein (Pup) is conjugated through its C-terminal residue to lysine side chains of substrates via an isopeptide bond. Pup proteins are small, ranging from 60 to 70 residues in length with Gly-Gly-Glu or Gly-Gly-Gln as C-terminal residues. Deamidation of the C-terminal glutamine of Pup by the deamidase of Pup (Dop) renders Pup competent for conjugation to substrates, which is then carried out by the Pup-ligase PafA
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the prokaryotic, ubiquitin-like protein (Pup) is conjugated through its C-terminal residue to lysine side chains of substrates via an isopeptide bond. Pup proteins are small, ranging from 60 to 70 residues in length with Gly-Gly-Glu or Gly-Gly-Gln as C-terminal residues. Deamidation of the C-terminal glutamine of Pup by the deamidase of Pup (Dop) renders Pup competent for conjugation to substrates, which is then carried out by the Pup-ligase PafA. The enzyme turns over one molecule of ATP to ADP per conjugation cycle, suggesting that PafA activates Pup by phosphorylation of its C-terminal glutamate. The formation of this intermediate occurs even in the absence of substrate and is shown to be the rate-limiting step of the PafA catalyzed reaction. After activation, the phosphorylated Pup and ADP remain bound to PafA, awaiting the nucleophilic attack of the substrate lysine, which finally results in the formation of the isopeptide bond. The C-terminal residue of the ligation-competent PupE features two carboxylates, the C-terminal alpha-carboxylate and the gamma-carboxylate of the glutamyl side chain, both of which could in principle be used for Pup-attachment. NMR-experiments using the pupylated proteasomal substrate PanB show that only the gamma-carboxylate is used to form the isopeptide bond between Pup and the target
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Pup is linked to substrate proteins by the formation of an isopeptide bond between the side-chain carboxylate of glutamate64 and a lysine side chain of the substrate protein
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the C-terminal 26 amino acid sequence of the prokaryotic ubiquitin-like protein (Pup) is the minimal ligase recognition motif in Mycobacterium tuberculosis. Specific hydrophobic residues within this sequence that are known to be important for the interactions of Pup with proteasomes are also critical for the activation of Pup by PafA
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the enzyme catalyzes a two-step reaction mechanism proceeding through a gamma-glutamyl phosphate-mixed anhydride intermediate that is formed on the C-terminal glutamate of the prokaryotic ubiquitin-like protein (Pup) before transfer of Pup to the substrate acceptor lysine. Phosphorylated [prokaryotic ubiquitin-like protein]-L-glutamate and ADP are retained on the enzyme in a stable fashion following activation
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
key enzyme in the Pup tagging (i.e. pupylation) system. Protein pupylation can be regulated at the enzyme level via an allosteric mechanism
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
PafA is an allosteric enzyme that binds its target substrates cooperatively. In contrast, binding of deamidated Pup (Pup-L-glutamate) by PafA follows simple kinetics and is, therefore, not involved in the allosteric transitions of PafA. Pup and target substrate binding by PafA are independent events
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
no activity with ATPgammaS
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycobacterium tuberculosis ATCC 25618 / H37Rv
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup product of the Dop reaction presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Mycolicibacterium smegmatis ATCC 700084 / mc2155
the Pup product of the Dop reaction presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Renibacterium salmoninarum ATCC 33209 / DSM 20767 / JCM 11484 / NBRC 15589 / NCIMB 2235
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Renibacterium salmoninarum ATCC 33209 / DSM 20767 / JCM 11484 / NBRC 15589 / NCIMB 2235
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Rhodococcus erythropolis PR4 / NBRC 100887
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Rhodococcus erythropolis PR4 / NBRC 100887
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Salinispora tropica ATCC BAA-916 / DSM 44818 / CNB-440
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Salinispora tropica ATCC BAA-916 / DSM 44818 / CNB-440
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Streptomyces coelicolor ATCC BAA-471 / A3(2) / M145
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
Streptomyces coelicolor ATCC BAA-471 / A3(2) / M145
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
-
-
-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-([prokaryotic ubiquitin-like protein]-gamma-L-glutamyl)-[protein]-L-lysine
the Pup variant used presents a glutamate at its C-terminus (PupE) and, as such, can be readily conjugated to target proteins by PafA, PupE is covalently attached to the protein via a GGL bridge, overview. The isopeptide bond to the lysine residue of the target protein occurs via the side-chain carboxylate
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-
?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-[[prokaryotic ubiquitin-like protein]-L-glutamate]-[[protein]-L-lysine]
pupylation targets the substrates for degradation by the proteasome
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?
ATP + [prokaryotic ubiquitin-like protein]-L-glutamate + [protein]-L-lysine
ADP + phosphate + N6-[[prokaryotic ubiquitin-like protein]-L-glutamate]-[[protein]-L-lysine]
pupylation targets the substrates for degradation by the proteasome
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?
additional information
?
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PafA substrate docking study, overview. PafA can use ammoniumions as targets for pupylation. The reaction catalyzed is in fact the conversion of PupE C-terminal glutamate into a glutamine, that is, PupE amidation. The docking of a small molecule like ammonium in the PafA active site cannot be supported by auxiliary interactions. PafA residue R207 stabilizes the interaction of the enzyme with PanB, the R207A mutant amidates PupE and the wild-type variant
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additional information
?
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Corynebacterium glutamicum ATCC 13032 / DSM 20300 / JCM 1318 / LMG 3730 / NCIMB 10025
PafA substrate docking study, overview. PafA can use ammoniumions as targets for pupylation. The reaction catalyzed is in fact the conversion of PupE C-terminal glutamate into a glutamine, that is, PupE amidation. The docking of a small molecule like ammonium in the PafA active site cannot be supported by auxiliary interactions. PafA residue R207 stabilizes the interaction of the enzyme with PanB, the R207A mutant amidates PupE and the wild-type variant
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additional information
?
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PafA can move Pup from one proteasome substrate, inositol 1-phosphate synthetase (Ino1), to two different proteins, malonyl coenzyme A (CoA)-acyl carrier protein transacylase (FabD) and lonely guy (Log). Mutagenesis of substrate FabD Lys173 to Ala results in reduced transfer of Pup from Pup-Ino1 in the transpupylation reaction. Pup-Ino1 is a better Pup donor than Pup-FabD, and transpupylation requires ATP. PafA needs depupylase activity to transfer Pup between proteins. Pupylation is a two-step reaction in which PafA uses ATP to phosphorylate a carboxylate on the C terminus of Pup, which is then attacked by the amino group of a lysine side chain
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additional information
?
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PafA can move Pup from one proteasome substrate, inositol 1-phosphate synthetase (Ino1), to two different proteins, malonyl coenzyme A (CoA)-acyl carrier protein transacylase (FabD) and lonely guy (Log). Mutagenesis of substrate FabD Lys173 to Ala results in reduced transfer of Pup from Pup-Ino1 in the transpupylation reaction. Pup-Ino1 is a better Pup donor than Pup-FabD, and transpupylation requires ATP. PafA needs depupylase activity to transfer Pup between proteins. Pupylation is a two-step reaction in which PafA uses ATP to phosphorylate a carboxylate on the C terminus of Pup, which is then attacked by the amino group of a lysine side chain
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additional information
?
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the prokaryotic ubiquitin-like protein remains intrinsically disordered when covalently attached to proteasomal target proteins. When linked to the proteasomal substrates FabD and PanB, Pup is unstructured and retains the ability to interact with its different binding partners. This suggests that it is not the conformation of Pup attached to these two substrates which determines their delivery to the proteasome, but the availability of the degradation complex and the depupylase. Structure analysis of Pup coupled to the PanB decamer, overview
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additional information
?
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the prokaryotic ubiquitin-like protein remains intrinsically disordered when covalently attached to proteasomal target proteins. When linked to the proteasomal substrates FabD and PanB, Pup is unstructured and retains the ability to interact with its different binding partners. This suggests that it is not the conformation of Pup attached to these two substrates which determines their delivery to the proteasome, but the availability of the degradation complex and the depupylase. Structure analysis of Pup coupled to the PanB decamer, overview
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Mycobacterium tuberculosis ATCC 25618 / H37Rv
the prokaryotic ubiquitin-like protein remains intrinsically disordered when covalently attached to proteasomal target proteins. When linked to the proteasomal substrates FabD and PanB, Pup is unstructured and retains the ability to interact with its different binding partners. This suggests that it is not the conformation of Pup attached to these two substrates which determines their delivery to the proteasome, but the availability of the degradation complex and the depupylase. Structure analysis of Pup coupled to the PanB decamer, overview
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Mycobacterium tuberculosis ATCC 25618 / H37Rv
PafA can move Pup from one proteasome substrate, inositol 1-phosphate synthetase (Ino1), to two different proteins, malonyl coenzyme A (CoA)-acyl carrier protein transacylase (FabD) and lonely guy (Log). Mutagenesis of substrate FabD Lys173 to Ala results in reduced transfer of Pup from Pup-Ino1 in the transpupylation reaction. Pup-Ino1 is a better Pup donor than Pup-FabD, and transpupylation requires ATP. PafA needs depupylase activity to transfer Pup between proteins. Pupylation is a two-step reaction in which PafA uses ATP to phosphorylate a carboxylate on the C terminus of Pup, which is then attacked by the amino group of a lysine side chain
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PafA can pupylate itself, extra-pupylation by PafA can also occur, mostly in the form of poly-pupylation. Unlike in the eukaryotic UPS where poly-ubiquitylation is seen, in vivo targets of the Pup-proteasome system are found almost exclusively to be monopupylated, except for PafA itself. Poly-pupylation occurs via pupylation of an already pupylated target, rather than by conjugation of pre-formed poly-Pup chains
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PafA binds IdeR with low affinity, compared to PanB, a model substrate whose affinity for PafA is about 10fold higher
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PafA can catalyze the poly-pupylation of itself, poly-pupylation of PafA. One common pupylation substrate is protein PanB. PafA self-pupylation is suppressed when PanB is added at high concentrations. Unlike the pattern of poly-pupylated PafA, only mono-pupylated PanB and a few di-pupylated PanB (Pup2-PanB) can be detected in the reaction system
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Mycolicibacterium smegmatis ATCC 700084 / mc2155
PafA can pupylate itself, extra-pupylation by PafA can also occur, mostly in the form of poly-pupylation. Unlike in the eukaryotic UPS where poly-ubiquitylation is seen, in vivo targets of the Pup-proteasome system are found almost exclusively to be monopupylated, except for PafA itself. Poly-pupylation occurs via pupylation of an already pupylated target, rather than by conjugation of pre-formed poly-Pup chains
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additional information
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Mycolicibacterium smegmatis ATCC 700084 / mc2155
PafA binds IdeR with low affinity, compared to PanB, a model substrate whose affinity for PafA is about 10fold higher
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additional information
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Mycolicibacterium smegmatis ATCC 700084 / mc2155
PafA can catalyze the poly-pupylation of itself, poly-pupylation of PafA. One common pupylation substrate is protein PanB. PafA self-pupylation is suppressed when PanB is added at high concentrations. Unlike the pattern of poly-pupylated PafA, only mono-pupylated PanB and a few di-pupylated PanB (Pup2-PanB) can be detected in the reaction system
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