Information on EC 2.4.2.43 - lipid IVA 4-amino-4-deoxy-L-arabinosyltransferase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Enterobacteriaceae

EC NUMBER
COMMENTARY
2.4.2.43
-
RECOMMENDED NAME
GeneOntology No.
lipid IVA 4-amino-4-deoxy-L-arabinosyltransferase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4-amino-4-deoxy-alpha-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate + alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid A = alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-[4-P-L-Ara4N]-lipid A + ditrans,octacis-undecaprenyl phosphate
show the reaction diagram
-
-
-
-
4-amino-4-deoxy-alpha-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate + alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA = 4'-alpha-L-Ara4N-alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA + ditrans,octacis-undecaprenyl phosphate
show the reaction diagram
-
-
-
-
4-amino-4-deoxy-alpha-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate + lipid IVA = lipid IIA + ditrans,octacis-undecaprenyl phosphate
show the reaction diagram
lipid IV(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose. Lipid II(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-alpha-D-glucopyranose
-
4-amino-4-deoxy-alpha-L-arabinopyranosyl ditrans,octacis-undecaprenyl phosphate + lipid IVA = lipid IIA + ditrans,octacis-undecaprenyl phosphate
show the reaction diagram
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
polymyxin resistance
-
SYSTEMATIC NAME
IUBMB Comments
4-amino-4-deoxy-alpha-L-2-aminoarabinopyranosyl ditrans,octacis-undecaprenyl phosphate:lipid IVA L-2-aminoarabinopyranosyltransferase
Integral membrane protein present in the inner membrane of certain Gram negative endobacteria. In strains that do not produce 3-deoxy-D-manno-octulosonic acid (Kdo), the enzyme adds a single arabinose unit to the 1-phosphate moiety of the tetra-acylated lipid A precursor, lipid IVA. In the presence of a Kdo disaccharide, the enzyme primarily adds an arabinose unit to the 4-phosphate of lipid A molecules. The Salmonella typhimurium enzyme can add arabinose units to both positions.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4-amino-4-deoxy-L-arabinose transferase
O52327
-
arnT
-
gene name
arnT
P76473
-
arnT
Escherichia coli K12
P76473
-
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
polymyxin-resistant mutant WD901 of strain K12
-
-
Manually annotated by BRENDA team
Escherichia coli K12
-
SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
P76473
LpxT-dependent lipid A modification is not restored in arnT or eptA mutants
malfunction
Escherichia coli K12
-
LpxT-dependent lipid A modification is not restored in arnT or eptA mutants
-
physiological function
-
ArnT confers resistance to the antibiotic polymyxin in Salmonella typhimurium and Escherichia coli through the modification of lipid A, a major component of the outer surface of Gram-negative bacteria. ArnT transfers a neutral aminoarabinose moiety onto the negative phosphate groups of lipid A, reducing the surface charge of the bacteria and preventing cationic peptides such as polymyxin from electrostatically recognizing and killing the bacteria. Only small amounts of ArnT are necessary to provide resistance against polymyxin to the bacterial cell
physiological function
P76473
PmrA controls synthesis of both ArnT and EptA
physiological function
Escherichia coli K12
-
PmrA controls synthesis of both ArnT and EptA
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
-
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
-
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
O52327
-
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
ArnT confers resistance to the antibiotic polymyxin in Salmonella typhimurium and Escherichia coli through the modification of lipid A, a major component of the outer surface of Gram-negative bacteria. ArnT transfers a neutral aminoarabinose moiety onto the negative phosphate groups of lipid A, reducing the surface charge of the bacteria and preventing cationic peptides such as polymyxin from electrostatically recognizing and killing the bacteria. Only small amounts of ArnT are necessary to provide resistance against polymyxin to the bacterial cell
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
the reaction is part of a enzymatic pathway by which Gram-negative bacteria acquire antibiotic resistance
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl undecaprenyl phosphate + lipid IV(A)
lipid II(A) + undecaprenyl phosphate
show the reaction diagram
-
ArnT adds a single 4-amino-4-deoxy-alpha-L-arabinose unit to the 1-phosphate moiety of the tetraacylated lipid A precursor, lipid IV(A), which lacks Kdo and forms lipid II (A). Transfer to lipid A occurs on the periplasmic side of the inner membrane. ArnT is a bifunctional glycosyltransferase in that it can incorporate two 4-amino-4-deoxy-alpha-L-arabinose units. Lipid IV(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose. Lipid II(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-alpha-D-glucopyranose, ArnT adds a single 4-amino-4-deoxy-alpha-L-arabinose unit to the 1-phosphate moiety of the tetraacylated lipid A precursor. Lipid IV(A), which lacks Kdo and forms lipid II (A)lipid IV(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose. Lipid II(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-alpha-D-glucopyranose
-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
O52327
-
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
ArnT confers resistance to the antibiotic polymyxin in Salmonella typhimurium and Escherichia coli through the modification of lipid A, a major component of the outer surface of Gram-negative bacteria. ArnT transfers a neutral aminoarabinose moiety onto the negative phosphate groups of lipid A, reducing the surface charge of the bacteria and preventing cationic peptides such as polymyxin from electrostatically recognizing and killing the bacteria. Only small amounts of ArnT are necessary to provide resistance against polymyxin to the bacterial cell
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl di-trans,poly-cis-undecaprenyl phosphate + lipid IVA
lipid IIA + di-trans,poly-cis-undecaprenyl phosphate
show the reaction diagram
-
the reaction is part of a enzymatic pathway by which Gram-negative bacteria acquire antibiotic resistance
-
-
?
4-amino-4-deoxy-alpha-L-arabinopyranosyl undecaprenyl phosphate + lipid IV(A)
lipid II(A) + undecaprenyl phosphate
show the reaction diagram
-
ArnT adds a single 4-amino-4-deoxy-alpha-L-arabinose unit to the 1-phosphate moiety of the tetraacylated lipid A precursor, lipid IV(A), which lacks Kdo and forms lipid II (A). Transfer to lipid A occurs on the periplasmic side of the inner membrane. ArnT is a bifunctional glycosyltransferase in that it can incorporate two 4-amino-4-deoxy-alpha-L-arabinose units. Lipid IV(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose. Lipid II(A) = 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-alpha-D-glucopyranose
-
-
?
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Triton X-100
-
transferase—transfer of the 4-amino-4-deoxy-alpha-L-arabinose unit from its putative isoprenoid carrier, undecaprenyl phosphate-4-amino-4-deoxy-alpha-L-arabinose, to the radiolabeled acceptor [4'-32P]lipid IV(A) is dependent upon the presence of the nonionic detergent Triton X-100, with maximal activity observed at 0.2% in the assay system
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Escherichia coli K12
-
-
-
Manually annotated by BRENDA team
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
the secondary structure of ArnT is not dependent on disulfide bridges, Salmonella typhimurium ArnT contains no disulfide bonds
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
purifiation of 6*His-tagged ArnT. HisLink nickel affinity chromatography coupled with anion exchange chromatography results in the best overall purifiation strategy for dodecylmaltoside-solubilized ArnT
O52327
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
overxpression of the L-Ara4N lipid A transferase (ArnT) behind a T7lac promoter
-
the 6*His-tagged ArnT gene-encoded plasmids are transformed into Escherichia coli NovaBlue (Novagen) cells for protein expression. The gene is knocked out of the chromosomal DNA of BL21 (BL21(DE3)DELTAArnT), leaving the plasmid-encoded ArnT to be the only ArnT expressed in the system
O52327
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C148A/C149A/C173S/C216S/C318S/C383S/C400S/C411S
-
when assayed for growth in the presence of polymyxin, the cysteine-free construct of ArnT supports growth at a level similar to that of the native protein
C148S/C149S/C173S/C216S/C318S/C383S/C400S/C411S
-
mutant enzyme is inactive
C173S/C216S/C318S/C383S/C400S/C411S
-
when assayed for growth in the presence of polymyxin, the cysteine-free construct of ArnT supports growth at 70% of the native protein
additional information
-
analysis of 31 point mutations within a putative periplasmic loop of the cysteine-free ArnT protein, carried out using an in vivo growth assay coupled with expression studies, identification of the first time specific critical residues within the bacterial transferase. These critical residues fall into two categories: those that disrupt initial protein folding or membrane localization and those that fail to confer in vivo resistance to polymyxin despite being expressed to the inner membrane