Information on EC 2.3.2.10 - UDP-N-acetylmuramoylpentapeptide-lysine N6-alanyltransferase

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The expected taxonomic range for this enzyme is: Weissella viridescens

EC NUMBER
COMMENTARY hide
2.3.2.10
-
RECOMMENDED NAME
GeneOntology No.
UDP-N-acetylmuramoylpentapeptide-lysine N6-alanyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-alanyl-tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine = tRNAAla + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminoacyl group transfer
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Peptidoglycan biosynthesis
-
-
peptidoglycan cross-bridge biosynthesis IV (Weissella viridescens)
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-
SYSTEMATIC NAME
IUBMB Comments
L-alanyl-tRNAAla:UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine N6-alanyltransferase
Also acts on L-seryl-tRNASer.
CAS REGISTRY NUMBER
COMMENTARY hide
37257-26-4
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain CIP 102810 T, formerly Lactobacillus viridescens ATCC 12706
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-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
glycyl-tRNAGly + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNAGly + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(glycyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
poor substrate
-
-
ir
L-alanyl-tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
L-alanyl-tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
-
-
?
L-cysteinyl-tRNACys + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNACys + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-cysteinyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
-
-
-
ir
L-seryl-tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNA + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-seryl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
-
-
?
L-seryl-tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNASer + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-seryl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
-
about half as effective as L-alanyl-tRNA
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ir
additional information
?
-
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
L-alanyl-tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
tRNAAla + UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-N6-(L-alanyl)-L-lysyl-D-alanyl-D-alanine
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
no activation by EDTA or metal ions
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
aminoacyl-tRNA analogue C
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-
-
aminoacyl-tRNA analogue C'
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-
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oxa-helixAla
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oxa-helixGly
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oxa-helixSer
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Ser-tRNA
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versus UDP-acetylmuramoylpentapeptide
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
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product inhibition
additional information
-
no inhibition by EDTA or metal ions
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.015 - 0.061
Ala-tRNAAla
0.0017
L-alanyl-tRNA
-
pH 7.5, 30°C
0.0002 - 0.165
UDP-acetylmuramoylpentapeptide
0.079
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
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pH 7.5, 30°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
26.7
L-alanyl-tRNA
Weissella viridescens
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pH 7.5, 30°C
0.183 - 11
L-alanyl-tRNAAla
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.248
Ser-tRNA
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versus UDP-acetylmuramoylpentapeptide
0.048
UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
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product inhibition
additional information
additional information
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-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0024
aminoacyl-tRNA analogue C
Weissella viridescens
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-
-
0.0041
aminoacyl-tRNA analogue C'
Weissella viridescens
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-
-
0.0014
oxa-helixAla
Weissella viridescens
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-
-
0.067
oxa-helixGly
Weissella viridescens
-
-
-
0.015
oxa-helixSer
Weissella viridescens
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-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
276
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purified enzyme
additional information
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relative activity of FemXWv for the transfer of L-Ala from helixAla to UDP-MurNAc-pentapeptide 100%; relative activity of FemXWv for the transfer of L-Ala from helixGly to UDP-MurNAc-pentapeptide 2.9%; relative activity of FemXWv for the transfer of L-Ala from helixSer to UDP-MurNAc-pentapeptide 31%
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 7.2
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L-alanyl-tRNA
7.2 - 7.6
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L-seryl-tRNA
additional information
-
broad maximum
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2 - 8.6
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L-alanyl-tRNA, about 80% of maximal activity at pH 5.2 and pH 8.6
6.1 - 8.6
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L-seryl-tRNA, about half-maximal activity at pH 6.1 and about 90% of maximal activity at pH 8.6
additional information
-
pK values of 5.5 and 9.3
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
inhibition assay
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
crystal structure analysis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 38000, crystal structure analysis
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystallized by vapor diffusion method
crystallized in the presence of UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine by vapor diffusion method
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutants from E. coli
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recombinant proteins using His-tag
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as intein-tagged fusion protein
expression of wild-type and mutants as His-tagged proteins in Escherichia coli, in vitro transcription of tRNAs
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wild type and mutant enzymes expressed as His-tag fusion proteins in Escherichia coli TOP10
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D109N
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site-directed mutagenesis, loss of 99% activity compared to the wild-type
E205Q
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site-directed mutagenesis, unaltered activity compared to the wild-type
E215Q
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site-directed mutagenesis, loss of 15% activity compared to the wild-type
E316Q
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site-directed mutagenesis, loss of 60% activity compared to the wild-type
E320Q
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site-directed mutagenesis, loss of 96% activity compared to the wild-type
F70L
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35% loss of activity
K36M
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activity complete abolished
K36R
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160-fold reduction of activity, 5.4-fold increased Km for UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
R211K
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17000-fold reduction of activity, no effect on Km for Ala-tRNA, Km for UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine higher than the highest sample concentration
Y103F
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no effect on activity
Y215F
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25-fold decrease of activity, increased Km for UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
Y215L
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insoluble protein
Y256F
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2.6-fold reduction of activity
F70L
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35% loss of activity
-
K36M
-
activity complete abolished
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R211K
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17000-fold reduction of activity, no effect on Km for Ala-tRNA, Km for UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine higher than the highest sample concentration
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Y103F
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no effect on activity
-
Y215F
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25-fold decrease of activity, increased Km for UDP-N-acetylmuramoyl-L-alanyl-D-glutamyl-L-lysyl-D-alanyl-D-alanine
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine