Information on EC 2.4.99.18 - dolichyl-diphosphooligosaccharide-protein glycotransferase

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The expected taxonomic range for this enzyme is: Eukaryota, Archaea, Bacteria

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EC NUMBER
COMMENTARY hide
2.4.99.18
-
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RECOMMENDED NAME
GeneOntology No.
dolichyl-diphosphooligosaccharide-protein glycotransferase
-
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dolichyl diphosphooligosaccharide + [protein]-L-asparagine = dolichyl diphosphate + a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to a protein L-asparagine
show the reaction diagram
-
-
-
-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexosyl group transfer
-
-
-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
protein N-glycosylation initial phase (eukaryotic)
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N-Glycan biosynthesis
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Various types of N-glycan biosynthesis
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
dolichyl-diphosphooligosaccharide:protein-L-asparagine N-beta-D-oligopolysaccharidotransferase
Occurs in eukaryotes that form a glycoprotein by the transfer of a glucosyl-mannosyl-glucosamine polysaccharide to the side-chain of an L-asparagine residue in the sequence -Asn-Xaa-Ser- or -Asn-Xaa-Thr- (Xaa not Pro) in nascent polypeptide chains. The basic oligosaccharide is the tetradecasaccharide Glc3Man9GlcNAc2 (for diagram {polysacc/Dol14}). However, smaller oligosaccharides derived from it and oligosaccharides with additional monosaccharide units attached may be involved. See ref [2] for a review of N-glycoproteins in eukaryotes. Man3GlcNAc2 seems to be common for all of the oligosaccharides involved with the terminal N-acetylglucosamine linked to the protein L-asparagine. Occurs on the cytosolic face of the endoplasmic reticulum. The dolichol involved normally has 14-21 isoprenoid units with two trans double-bonds at the omega end, and the rest of the double-bonds in cis form.
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CAS REGISTRY NUMBER
COMMENTARY hide
75302-32-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
NBRC 100126, three AglB paralogues
UniProt
Manually annotated by BRENDA team
calf
-
-
Manually annotated by BRENDA team
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B9LMS1
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
Q12VP3
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
no activity in Aeropyrum pernix
-
-
-
Manually annotated by BRENDA team
no activity in Methanopyrus kandleri
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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A1RRM7
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
syn. Pyrococcus kodakaraensis
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
biantennary Man5GlcNAc2 dolichyl disphosphate + VSG221 coat glycoprotein
dolichyl disphosphate + ?
show the reaction diagram
-
selective transfer of TbSTT3A to acidic to neutral regions of polypeptides, responsible for all paucimannose and complex N-glycans in Trypanosoma brucei glycoproteins
-
-
?
biantennary Man5GlcNAc2 dolichyl disphosphate + VSG221 coat glycoprotein
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine
show the reaction diagram
-
selective transfer of TbSTT3A to acidic to neutral regions of polypeptides, responsible for all paucimannose and complex N-glycans in Trypanosoma brucei glycoproteins
-
-
?
Campylobacter jejuni heptasaccharide bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
alpha-1,3-linked sugar, in vivo in Escherichia coli
-
-
?
crude lipid-linked oligosaccharide donors from Campylobacter jejuni + carboxytetramethylrhodamine-Ala-Asp-Gln-Asn-Ala-Thr-Tyr-Lys
dolichyl disphosphate + carboxytetramethylrhodamine-Ala-(oligosaccharidyl) Asp-Gln-Asp-Ala-Thr-Tyr
show the reaction diagram
dolichyl diphosphate-di-N-acetylchitobiose + Tyr-Asn-Leu-Thr-Ser-Val
?
show the reaction diagram
dolichyl diphosphochitobiose-(mannosyl)9-(glucosyl)3 + synthetic hexapeptide
?
show the reaction diagram
dolichyl diphosphooligosaccharide + asparagine-asparagine-threonine-NH2 acceptor peptide
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine
show the reaction diagram
-
-
-
?
dolichyl diphosphooligosaccharide + carboxypeptidase Y (CPY)
dolichyl disphosphate + oligosaccharidyl-(carboxypeptidase Y)
show the reaction diagram
-
free enzyme
-
-
?
dolichyl diphosphooligosaccharide + N2-acetyl-L-asparaginyl-4-benzoyl-L-phenylalanyl-L-tyrosinamide
dolichyl diphosphate + ?
show the reaction diagram
dolichyl diphosphooligosaccharide + protein L-asparagine
dolichyl diphosphate + a glycoprotein with the oligosaccharide chain attached by N-glycosyl linkage to protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + protein L-asparagine
dolichyl diphosphate + glycoprotein with the oligosaccharide chain attached by N-glycosyl-linkage to protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + protein L-asparagine
dolichyl diphosphate + oligosaccaharidyl-L-Asn protein
show the reaction diagram
dolichyl diphosphooligosaccharide + protein L-asparagine
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + synthetic tripeptides
?
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to a protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + [protein]-L-asparagine-N-oligosaccharide
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + [protein]-L-asparagine-oligosaccharide
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
show the reaction diagram
dolichyl monophosphate GlcNAc-Glc-2,3-diNAcA + [protein]-L-asparagine
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose + Arg-Asn-Gly-Thr-Ala-Val-methylester
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose + N-benzoyl-Asn-Gly-Thr-NHCH3
?
show the reaction diagram
dolichyl-diphosphochitobiose + Tyr-Asn-Leu-Thr-Ser-Val
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose-Man1 + Tyr-Asn-Leu-Thr-Ser-Val
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose-Man7Glc3 + protein-L-asparagine
?
show the reaction diagram
dolichyl-diphosphochitobiose-Man9 + Nalpha-Ac-Asn-Tyr-Thr-NH2
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose-Man9Glc3 + alpha-Ac-Asn-Tyr-Thr-NH2
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose-Man9Glc3 + Nalpha-Ac-Asn-Lys(Nepsilon-p-azidobenzoyl)-Thr-NH2
?
show the reaction diagram
-
-
-
-
?
dolichyl-diphosphochitobiose-Man9Glc3 + protein-L-asparagine
?
show the reaction diagram
dolichyl-diphosphochitobiose-Man9Glc3 + Tyr-Asn-Leu-Thr-Ser-Val
?
show the reaction diagram
-
-
-
-
?
Escherichia coli group 1 K30 capsule antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli
-
-
?
Escherichia coli O16 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
alpha-1,6-linked Glc with GlcNAc, in vivo in Escherichia coli
-
-
?
Escherichia coli O2 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
rhamnose beta-1,4-linked sugar, in vivo in Escherichia coli
-
-
?
Escherichia coli O7 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
beta-1,3-linked sugar, in vivo in Escherichia coli
-
-
?
Escherichia coli O86 antigen bound to farnesyl diphosphate + pilin
farnesyl diphosphate + glycosylated pilin
show the reaction diagram
-
pentasaccharide attached to other lipid carrier, in vitro, 500 mM Tris-HCl containing 1 M sucrose and 10 mM MnCl2, pH7.5, 30°C
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DFNAT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DFNVT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DQNAT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DVNAS-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DVNAT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-DVNVT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
GalNAc-N,N'-diacetyl-bacillosamine-PP-undecaprenyl + acetyl-EVNAT-(4-nitrophenylalanine)-NH2
?
show the reaction diagram
-
-
-
-
?
Glc3Man9GlcNAc2 dolichyl diphosphate + protein L-asparagine
?
show the reaction diagram
-
-
-
-
?
Glc3Man9GlcNAc2 dolichyl disphosphate + GAYNSTSV
dolichyl disphosphate + GAY-(Glc3Man9GlcNAc2)NSTSV
show the reaction diagram
-
microsomes as enzyme source, STT3-1, increase in activity from 51 to 82% when the yeast SST3 expression is turned off, when yeast SST3 is active, only slight change in activity to 103%
-
-
?
lipid-linked oligosaccharide + unfolded nascent polypeptide chain
?
show the reaction diagram
Man5GlcNAc2 dolichyl diphosphate + GAYNSTSV
dolichyl diphosphate + GAY-(Man5GlcNAc2)NSTSV
show the reaction diagram
-
microsomes as enzyme source, STT3-1, increase in activity from 34 to 109% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 115%
-
-
?
Man6GlcNAc2 dolichyl diphosphate + GAYNSTSV
dolichyl diphosphate + GAY-(Man6GlcNAc2)NSTSV
show the reaction diagram
-
microsomes as enzyme source, STT3-1, increase in activity from 46 to 151% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 173%
-
-
?
Man9GlcNAc2 dolichyl disphosphate + GAYNSTSV
dolichyl disphosphate + GAY-(Man9GlcNAc2)NSTSV
show the reaction diagram
-
microsomes as enzyme source, STT3-1, increase in activity from 54 to 172% when the yeast SST3 expression is turned off, when yeast SST3 is active, increase in activity to 202%
-
-
?
Man9GlcNAc2PP dolichyl diphosphate + protein L-asparagine
?
show the reaction diagram
-
-
-
-
?
N-acetyl-D-galactosaminyl-alpha-(1->3)-N,N'-diacetyl-alpha-D-bacillosaminyl-diphospho-tritrans,heptacis-undecaprenol + KDFNVSKA
tritrans,heptacis-undecaprenyl diphosphate + Lys-Asp-Phe-N4-[N-acetyl-D-galactosaminyl-alpha-(1->3)-N,N'-diacetyl-beta-D-bacillosaminyl]-Asn-Val-Ser-Lys-Ala
show the reaction diagram
-
-
-
-
?
N-acetyl-D-glucosamine + glycoprotein bound to polysaccharide cell wall
?
show the reaction diagram
-
glycoproteins as substrates are P15703, O13547, P53301, P32623, P28319, P38248, P22146, Q03655, Q08193, P38616
-
-
?
pentaprenyl diphospho-N-acetyl-D-galactosamine + [protein]-L-asparagine
pentaprenyl diphosphate + [protein]-N-(N-acetyl-D-galactosaminyl)-L-asparagine
show the reaction diagram
PglB can utilize shorter polyisoprenol (pentaprenol) as the lipid carrier, albeit with reduced efficiency
-
-
?
peptidoglycan bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
transfer of subunit of the peptidoglycan petapeptide, no transfer of the complete peptidoglycan is observed, in vivo in Salmonella enterica
-
-
?
Pseudomonas aeruginosa O11 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
FucNac, in vivo in Escherichia coli
-
-
?
Salmonella enterica O antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli
-
-
?
Salmonella typhimurium LT2 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli, in vivo in Salmonella enterica serovar Typhimurium LT2 strain
-
-
?
triantennary Man9GlcNAc2 dolichyl disphosphate + VSG221 coat glycoprotein
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine
show the reaction diagram
-
selective transfer of TbSTT3B to neutral to basic regions of polypeptides responsible for most or all oligomannose N-glycans in Trypanosoma brucei glycoproteins
-
-
?
undecaprenyl diphospho-N-acetyl-D-galactosamine + [protein]-L-asparagine
undecaprenyl diphosphate + [protein]-N-(N-acetyl-D-galactosaminyl)-L-asparagine
show the reaction diagram
PglB is solely responsible for the oligosaccharyltransferase activity. PglB can transfer a monosaccharide, e.g. GalNAc to a peptide acceptor. PglB exhibits relaxed sugar substrate specificity
-
-
?
additional information
?
-
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Campylobacter jejuni heptasaccharide bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
alpha-1,3-linked sugar, in vivo in Escherichia coli
-
-
?
dolichyl diphosphooligosaccharide + protein L-asparagine
dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + a glycoprotein with the oligosaccharide chain attached by N-beta-D-glycosyl linkage to a protein L-asparagine
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + [protein]-L-asparagine-N-oligosaccharide
show the reaction diagram
dolichyl diphosphooligosaccharide + [protein]-L-asparagine
dolichyl diphosphate + [protein]-N-oligosaccharidyl-L-asparagine
show the reaction diagram
Escherichia coli group 1 K30 capsule antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli
-
-
?
Escherichia coli O16 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
alpha-1,6-linked Glc with GlcNAc, in vivo in Escherichia coli
-
-
?
Escherichia coli O2 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
rhamnose beta-1,4-linked sugar, in vivo in Escherichia coli
-
-
?
Escherichia coli O7 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
beta-1,3-linked sugar, in vivo in Escherichia coli
-
-
?
Escherichia coli O86 antigen bound to farnesyl diphosphate + pilin
farnesyl diphosphate + glycosylated pilin
show the reaction diagram
-
pentasaccharide attached to other lipid carrier, in vitro, 500 mM Tris-HCl containing 1 M sucrose and 10 mM MnCl2, pH7.5, 30°C
-
-
?
lipid-linked oligosaccharide + unfolded nascent polypeptide chain
?
show the reaction diagram
N-acetyl-D-glucosamine + glycoprotein bound to polysaccharide cell wall
?
show the reaction diagram
-
glycoproteins as substrates are P15703, O13547, P53301, P32623, P28319, P38248, P22146, Q03655, Q08193, P38616
-
-
?
peptidoglycan bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
transfer of subunit of the peptidoglycan petapeptide, no transfer of the complete peptidoglycan is observed, in vivo in Salmonella enterica
-
-
?
Pseudomonas aeruginosa O11 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
FucNac, in vivo in Escherichia coli
-
-
?
Salmonella enterica O antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli
-
-
?
Salmonella typhimurium LT2 antigen bound to undecaprenyl diphosphate + pilin
undecaprenyl diphosphate + glycosylated pilin
show the reaction diagram
-
in vivo in Escherichia coli, in vivo in Salmonella enterica serovar Typhimurium LT2 strain
-
-
?
additional information
?
-
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Arg-Asn-Gly-(Gly-allyl)-Ala-Val-methyl ester
-
-
Arg-Asn-Gly-(Gly-allylepoxide)-Ala-Val-methyl ester
-
-
Arg-Asn-Gly-(Gly-epoxyethylglycine)-Ala-Val-methylester
-
-
Arg-Asn-Gly-(Gly-vinyl)-Ala-Val-methyl ester
-
-
Digitonin
-
above 0.1%
L-threoninamide, (2S)-2-(benzoylamino)-4-[(phenylmethyl)amino]butanoylglycyl-L-threonyl-L-valyl
-
-
L-threoninamide, (2S)-2-(benzoylamino)-4-[[(4-methoxyphenyl)methyl]amino]butanoylglycyl-L-threonyl-L-valyl
-
-
L-threoninamide, (2S)-2-(benzoylamino)-4-[[(4-nitrophenyl)methyl]amino]butanoylglycyl-L-threonyl-L-valyl
-
-
L-Threoninamide, (2S)-2-[(6-mercapto-1-oxohexyl)amino]-4-[(2-naphthalenylmethyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-2-[(6-mercapto-1-oxohexyl)amino]-4-[(2-naphthalenylmethyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-N-(2-phenylethyl)-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-2-[(6-mercapto-1-oxohexyl)amino]-4-[(2-naphthalenylmethyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-N-[2-(4-nitrophenyl)ethyl]-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-4-(decylamino)-2-[(6-mercapto-1-oxohexyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-, cyclic (1-2)-thioether
-
-
L-threoninamide, (2S)-4-amino-2-[(6-mercapto-1-oxohexyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-4-amino-2-[(6-mercapto-1-oxohexyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-N-(2-phenylethyl)-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-4-amino-2-[(6-mercapto-1-oxohexyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-N-(4-phenylbutyl)-, cyclic (1-2)-thioether
-
-
L-Threoninamide, (2S)-4-amino-2-[(6-mercapto-1-oxohexyl)amino]butanoyl-L-cysteinyl-L-threonyl-L-valyl-N-[2-(4-nitrophenyl)ethyl]-, cyclic (1-2)-thioether
-
-
N-benzoyl-Asn-Leu-Thr
-
-
N-dinitrobenzoyl-Arg-Asn-Ala-epoxyethylglycine-Ala-Val
-
Asn residue of the inhibitor is glycosylated during inactivation
N-octanoyl-Asn-Leu-Thr
-
-
N-t-butoxycarbonyl-Asn-Leu-Thr
-
-
octylglucoside
-
concentration 1%
RNAi
-
silencing of TbSTT3A
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Digitonin
-
purified enzyme requires 0.1% digitonin for maximal activity
phosphatidylcholine
-
optimal concentration: 2 mM
additional information
-
DAD1 might provide structural and functional integrity for the OST complex
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2
Ac-Asn-Ala-Thr-NH2
-
-
1
Ac-Asn-Leu-Thr-NH2
-
-
-
0.08 - 0.143
acetyl-Asn-Ala-Thr
0.278
acetyl-Asn-Lys-Thr
-
-
0.00087
acetyl-DFNAT-(4-nitrophenylalanine)-NH2
-
-
0.00122
acetyl-DFNVT-(4-nitrophenylalanine)-NH2
-
-
0.0008
acetyl-DQNAT-(4-nitrophenylalanine)-NH2
-
-
0.003
acetyl-DVNAS-(4-nitrophenylalanine)-NH2
-
-
0.00106
acetyl-DVNAT-(4-nitrophenylalanine)-NH2
-
-
0.00144
acetyl-DVNVT-(4-nitrophenylalanine)-NH2
-
-
0.0225
acetyl-EVNAT-(4-nitrophenylalanine)-NH2
-
-
0.3 - 0.358
Ala-Leu-Gln-Asn-Ala-Thr-Arg
0.3
Ala-Leu-Glu-Asn-Ala-Thr-Arg-NH2
-
-
-
0.56 - 2.09
Asn-Ala-Thr
3.3
Asn-Asp-Thr
-
-
0.25
benzoyl-Asn-Leu-Thr-N-methyl-threonine
-
-
0.072 - 0.23
diphenyl-Ala-Leu-Glu-Asn-Ala-Thr-Arg-NH2
0.23
diphenyl-AlaLeu-Glu-Asn-Ala-Thr-Arg-NH2
-
-
0.0012
dolichyl diphosphochitobiose
-
-
0.0005
dolichyl diphosphochitobiose-(Man)9-(Glc)3
-
-
0.0315
dolichyl-diphosphochitobiose-(Man)9
-
-
0.021
dolichyl-diphosphochitobiose-(Man)9-(Glc)3
-
-
12.5
N-benzoyl-Asn-Gly-D-allo-Thr-NHCH3
-
-
4.3
N-benzoyl-Asn-Gly-L-threo-beta-hydroxynorvaline-NHCH3
-
-
0.6
N-benzoyl-Asn-Gly-Ser-NHCH3
-
-
0.16
N-benzoyl-Asn-Gly-Thr-NHCH3
-
-
0.00015
Nalpha-Ac-Asn-Tyr-Thr-NH2
-
-
0.05 - 0.6
Tyr-Asn-Leu-Thr-Ser-Val
0.08 - 0.127
Tyr-Gln-Ser-Asn-Ser-Thr-Met
additional information
additional information
-
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 7.5
-
-
7.4
-
assay at
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
-
pH 6.0: about 30% of maximal activity, pH 8.0: about 20% of maximal activity
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24
-
assay at
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
lymphoma cell line CH12-LX
Manually annotated by BRENDA team
-
about 95% purity
Manually annotated by BRENDA team
-
hepatoma cell line
Manually annotated by BRENDA team
additional information
-
TbSTT3A transcribed in the bloodstream nor procyclic form; TbSTT3B transcribed in the bloodstream nor procyclic form; TbSTT3C not transcribed in the bloodstream nor procyclic form
Manually annotated by BRENDA team
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
oligosaccharyltransferase forms a binary complex with ribosomes. Reconstitution of a binary complex containing oligosaccharyltransferase and ribosomes and its electron microscopic images. Oligosaccharyltransferase, the Sec61 complex, and ribosomes form a ternary complex in vitro
Manually annotated by BRENDA team
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PDB
SCOP
CATH
ORGANISM
UNIPROT
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6000
-
x * 62000-64000, Ost1p, + x * 6000, Stt3p, + x * 47000, Wbp1p, + x * 34000, Ost3p, + x * 32000, Ost6p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p
31500
MALDI-TOF result in accordance with calculated results for histidine-tagged protein subunit Stt3p
32000
-
x * 62000-64000, Ost1p, + x * 6000, Stt3p, + x * 47000, Wbp1p, + x * 34000, Ost3p, + x * 32000, Ost6p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p
45000
-
x * 78000, Stt3p, + x * 64000, Ost1p, + x * 45000, Wbp1p, + x * 34000, Ost3p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p, the enzyme is composed of equimolar amounts of eight subunits, the enzyme is composed of the following three subcomplexes: 1. Stt3p-Ost4p-Ast3p, 2. Swp1p-Wbp1p-Ost2p0, 3. Ost1p-Aso5p, SDS-PAGE
47000
-
x * 62000-64000, Ost1p, + x * 6000, Stt3p, + x * 47000, Wbp1p, + x * 34000, Ost3p, + x * 32000, Ost6p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p
48900
-
x * 48900, OST48, + x * 68700, ribophorin I, + x * 69300, ribophorin II + ?, calculation from nucleotide sequence
60000
-
x * 60000, SDS-PAGE
63000
-
x * 66000 + x * 63000 + x * 48000, SDS-PAGE
64000
-
x * 78000, Stt3p, + x * 64000, Ost1p, + x * 45000, Wbp1p, + x * 34000, Ost3p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p, the enzyme is composed of equimolar amounts of eight subunits, the enzyme is composed of the following three subcomplexes: 1. Stt3p-Ost4p-Ast3p, 2. Swp1p-Wbp1p-Ost2p0, 3. Ost1p-Aso5p, SDS-PAGE
66000
-
x * 66000 + x * 63000 + x * 48000, SDS-PAGE
68700
-
x * 48900, OST48, + x * 68700, ribophorin I, + x * 69300, ribophorin II + ?, calculation from nucleotide sequence
69300
-
x * 48900, OST48, + x * 68700, ribophorin I, + x * 69300, ribophorin II + ?, calculation from nucleotide sequence
78000
-
x * 78000, Stt3p, + x * 64000, Ost1p, + x * 45000, Wbp1p, + x * 34000, Ost3p, + x * 30000, Swp1p, + x * 16000, Ost2p, + x * 9500, Ost5p, + x * 3600, Ost4p, the enzyme is composed of equimolar amounts of eight subunits, the enzyme is composed of the following three subcomplexes: 1. Stt3p-Ost4p-Ast3p, 2. Swp1p-Wbp1p-Ost2p0, 3. Ost1p-Aso5p, SDS-PAGE
92000
-
recombinant monomeric Leishmania major SST3 enzyme with FLAG-tag, calculated and confirmed by SDS-PAGE
105000
-
2 * 105000, SDS-PAGE
180000
-
non-denaturing PAGE; recombinant Leishmania major SST3 with FLAG-tag, BN-PAGE probed with anti-FLAG antiserum
230000
-
BN-PAGE
480000
500000
550000
-
non-denaturing PAGE
600000
-
complex OSTC(II), native PAGE
700000
-
complex OSTC(III), native PAGE
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SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heptamer
monomer
-
1 * 180000
nonamer
-
the enzyme consists of nine subunits. Five of them, Wbp1, Swp1, Stt3, Ost1, and Ost2, are essential for viability of the cell, whereas Ost4, Ost5, Ost3, and Ost6 are not essential but are required for maximal OST activity: Wbp1 (49392 Da), Swp1 (31653 Da), Stt3 (81529 Da), Ost1 (54072 Da), Ost2 (14698 Da), Ost3, Ost4, Ost5, and Ost6, calculated from sequence
octamer
-
eight polypeptides assemble into a heterooctameric yeast OST complex composed of one copy each of Ost1p, Ost2p, Ost3p or Ost6p, Ost4p, Ost5p, Wbp1p, Swp1p, and Stt3p
additional information
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
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Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
C-terminal globular domain of the of the smallest AglB, purified recombinant wild-type and selenomethionine-labeled AgBs, sitting and hanging drop vapor diffusion methods, crystals grow from a hanging drop with a 1:1 volume ratio of the protein stock solution containing 20 mg/mL protein in 20 mM Tris buffer, pH 8.0, and the reservoir solution containing 0.1 M MES buffer, pH 6.5, and 12.5% PEG 3350, at 20°C, soaking of crystals in reservoir solution with 20% ethylene glycol added for cryoprotection, X-ray diffraction structure determination and analysis at 1.75-1.94 A resolution
sitting drop vapor diffusion method, crystal structure of the C-terminal globular domain of the AglB paralog, AglB-Long, at 1.9 A resolutions
2.8 A resolution crystal structure of the C-terminal globular domain, hanging drop method; hanging drop crystallization with 10 mM Tris-HCl, pH 8.0, and reservoir solution (0.1 M sodium cacodylate, pH 6.5, 18% polyethylene glycol 8000, 0.2 M calcium acetate, 20°C, cryoprotection with 0.1 M MES, pH 6.5, polyethylene glycol 8000, and 0.2 M calcium acetate)
2.7 A resolution crystal structure of the C-terminal soluble domain of Pyrococcus STT3
-
sitting-drop vapour diffusion method
-
12 A resolution cryoelectron microscopy structure
-
sitting drop vapor diffusion method, dolichyl-diphosphooligosaccharide-protein glycosyltransferase subunit OST6
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
3 h, stable
50
-
enzyme in endoplasmic reticulum preparations is stable below
54
-
absolute melting temperature of large soluble domain of the subunit Nlt1p
60
-
inactivation by heating of endoplasmic reticulum preparations
65
-
absolute melting temperature of large soluble domain of the subunit Swp1p
73
-
absolute melting temperature of large soluble domain of the subunit Wbp1p
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
DAD1 might provide structural and functional integrity for the OST complex
-
glycerol, 25%, stabilizes
-
phosphatidylcholine stabilizes detergent-solubilized enzyme
-
solubilization with detergents inactivates
-
strongly stabilized by addition of phospholipids upon detergent solubilization, phosphatidylcholine is twice as effective as phosphatidylethanolamine
-
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, Nonidet-solubilized crude enzyme, 25% glycerol, 0.01% 2-mercaptoethanol, 18% loss of activity after 1 month and 87% after 5 months
-
-70°C, Nonidet-solubilized crude enzyme, 25% glycerol, 0.01% 2-mercaptoethanol, retains 90% of original activity after 1 month and 55% after 5 months
-
-85°C, membrane preparation, 30% glycerol, several months
-
4°C, less than 20% loss of activity within 1 week
-
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity purification of a tagged Stt3 protein (component of the oligosaccharyltransferase complex)
C-terminal domain of Stt3p, wild-type and mutant D518E; dissolution in denaturing buffer (6 M guanidine hydrochloride, 500 mM Na Cl, 25 mM imidazole, 20 mM phosphate buffer, pH 7.4), cenrtrifugation, supernatant loaded onto nickel-NTA column for affinity chromatography with 20 mM phosphate buffer, pH 6.5, SDS-PAGE
cells centrifuged, resuspended, and washed in 20 mM phosphate buffer, pH 7.2, with 0.3 M Na Cl, cells disrupted and centrifuged, membranes separated by ultracentrifugation of supernatant, resuspended in buffer with 2% Elugent and 25 mM imidazole buffer, proteins solubilized by tumbling, centrifuged, supernatant removed, Elugenat reduced to 1% or 0.5% n-dodecyl-beta-D-maltopyranoside, loaded onto Ni-nitrilotriacetic acid agarose column, elution with buffer containing 250 mM imidazole
-
Ni-NTA resin column chromatography
-
partial
purification of an affinity-tagged version of the enzyme complex from a membrane protein fraction
-
recombinant N-terminally His6-tagged wild-type and selenomethionine-labeled AglB from Escherichia coli strain BL21 by nickel affinity chromatography and gel filtration, followed by anion exchange chromatography
sonication, centrifugation, supernatant absorbed to glutathione-Sepharose 4B resin, elution, concentration, reductive methylation, gel filtration with Superdex75 column, followed by anion exchange chromatography with a Resource Q column
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; expression in Escherichia coli
-
cloning of DNA encoding the full-length AfAglB-S1 gene, expression of N-terminally His6-tagged wild-type and selenomethionine-labeled AglB in Escherichia coli strains BL21(DE3) and C43(DE3), respectively
cloning of OST48, ribophorin I, ribophorin II and expression in COS-1 cells
-
creation of a yeast strain in which the essential 64000 Da glycoprotein Nlt1p subunit of the oligosaccharyl transferase is modified by the addition of a 22-residue carboxy-terminal affinity tag, the tag includes both an 8-residue FLAG epitope and a 6-residue histidine motif
-
despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloarcula marismortui can readily replace their counterpart from Haloferax volcanii when introduced into Haloferax volcanii cells deleted of aglB
-
despite processing distinct lipid-linked glycans in their native hosts, AglB from Halobacterium salinarum can readily replace their counterpart from Haloferax volcanii when introduced into Haloferax volcanii cells deleted of aglB
-
despite processing distinct lipid-linked glycans in their native hosts, AglB from Haloferax mediterranei can readily replace their counterpart from Haloferax volcanii when introduced into Haloferax volcanii cells deleted of aglB
-
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells
-
expression in Escherichia coli
functionally expressed in Escherichia coli
-
Helicobacter species contain two unrelated pglB genes (pglB1 and pglB2), neither of which is located within a larger locus involved in protein glycosylation. In complementation experiments, the Helicobacter pullorum PglB1 protein, but not PglB2, is able to transfer Campylobacter jejuni N-linked glycan onto an acceptor protein in Escherichia coli
Lm STT3 paralogues LmSTT3A, LmSTT3B, LmSTT3C, or LmSTT3D are expressed individually in Saccharomyces cerevisiae. Three complemented the yeast stt3 deletion. LmSTT3D expression suppresses deletions of other essential OTase subunits (Wbp1p, Ost1p, Ost2p, and Swp1p). Leishmania major STT3 paralogues are active enzymes that do not incorporate into the yeast OTase complex, but instead form dimers
-
OST complex is composed of ribophorin I (Ost1p), ribophorin II (Swp1p), OST48 (Wbp1p), DAD1 (Ost2p), and an STT3 homologue
-
PCR-amplification fused to GST, expressed in Escherichia coli BL21 (DE3)pLysS; the expression plasmid encoding the C-terminal soluble domain (residues 428–713) is constructed by inserting a PCR product from genomic DNA (ATCC700819D) into the pGEX-6P-1 vector (GE Healthcare). The GST-fused sPglB protein is expressed by the addition of isopropyl-beta-D-thiogalactopyranoside at 37°C in the Escherichia coli BL21(DE3)pLysS strain. A structure-aided multiple sequence alignment of the STT3/PglB/AglB protein family reveals three types of oligosaccharyltransferases catalytic centers. This novel classification will provide a useful framework for understanding the enzymatic properties of the oligosaccharyltransferases enzymes from eukarya, archaea, and bacteria
PCR-amplification, expression of histidine-tagged enzyme in Escherichia coli and Salmonella enterica strains
-
PCR-amplification, expression of the pure C-terminal domain of the subunit Stt3p in Escherichia coli BL21 (DE3); subcloning, overexpression, and a method of production of the pure C-terminal domain of Stt3p at 60-70 mg/l in Escherichia coli
PCR-amplification, FLAG-tag, expression in Saccharomyces cerevisiae; STT3 from Leishmania is able to complement stt3 deficiency in yeast during vegetative growth, but only poorly during sporulation. The Leishmania STT3 homolog is functional mainly as a free enzyme with a broad specificity for the glycosyl donor. When incorporated into the OST complex, it accepts the common Glc3Man9GlcNAc2-PP-Dol donor. Three out of the four STT3 paralogs are able to functionally complement not only an stt3 mutant but also ost1, ost2, wbp1, or swp1 mutants. STT3 from Leishmania is not incorporated into the complex and most likely functions as a monomeric enzyme
-
PCR-amplification, TbSTT3A expression in Saccharomyces cerevisiae does not replace yeast SST3; PCR-amplification, TbSTT3B expression in Saccharomyces cerevisiae can replace yeast SST3, mild temperature-sensitive growth at 37°C, wild-type growth at 23°C; PCR-amplification, TbSTT3C expression in Saccharomyces cerevisiae can replace yeast SST3, severe growth defects at 23°C
-
PglB is overexpressed in Escherichia coli C43(DE3) at a level of 1 mg/L cell cultures
plasmids with subunit genes introduced into mutants lacking both subunits
-
preparation of a membrane fraction from Escherichia coli in which PglB has been overexpressed
-
replacement of Saccharomyces cerevisiae Stt3p by the Trypanosoma cruzi homologue
-
the C-terminal domain of the STT3 protein of Pyrococcus furiosus is expressed in Escherichia coli cell BL21
-
the large soluble domains of the subunits Nlt1p, Swp1p and Wbp1p are expressed in a baculovirus-infected insect cell system
-
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
induced by the addition of copper to media
-
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D520A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
D520N
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
D526A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
E521A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
E521Q
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
I522A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
I535A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
K531A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows similar OST activity compared to the wild-type enzyme
K531D
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
K531E
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
K531R
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
M524A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
N528A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
T518A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
T523A
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
D520A
-
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
-
D520N
-
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
-
D526A
-
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant cannot be expressed in Escherichia coli
-
E521Q
-
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
-
T523A
-
site-directed mutagensis of the AglB-S1 subunit of OST, the mutant shows reduced OST activity compared to the wild-type enzyme
-
D54A
XXD motif mutation, strongly reduced activity compared to wild-type
I571A
MI motif mutation, strongly reduced activity compared to wild-type
L570A
MI motif mutation, similar activity as wild-type