Information on EC 2.4.1.227 - undecaprenyldiphospho-muramoylpentapeptide beta-N-acetylglucosaminyltransferase

peptidoglycan glycosyltransferase

proteins, gene murG

-

translocase II

-

UDP-acetylglucosamine-acetylmuramoylpentapeptide pyrophospholipid acetylglucosaminyltransferase

-

UDP-N-acetyl-alpha-D-glucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol = UDP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

transfer of alpha-N-acetylglucosamine

peptidoglycan biosynthesis

BRENDA

KEGG

Peptidoglycan biosynthesis

MetaCyc

peptidoglycan biosynthesis I (meso-diaminopimelate containing), peptidoglycan biosynthesis II (staphylococci), peptidoglycan biosynthesis III (mycobacteria), peptidoglycan biosynthesis IV (Enterococcus faecium), peptidoglycan biosynthesis V (beta-lactam resistance)

UDP-N-acetyl-D-glucosamine:N-acetyl-alpha-D-muramyl(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol 4-beta-N-acetylglucosaminlytransferase

The enzyme also works when the lysine residue is replaced by meso-2,6-diaminoheptanedioate (meso-2,6-diaminopimelate, A2pm) combined with adjacent residues through its L-centre, as it is in Gram-negative and some Gram-positive organisms. The undecaprenol involved is ditrans,octacis-undecaprenol (for definitions, {iupac/misc/prenol#p6::click here}).

60976-26-3

-

dUDP-N-acetyl-alpha-D-glucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

?

P17443

-

488977

-

?

UDP-N-acetylglucosamine + biotin-labelled lipid I analogue

?

-

-

?

UDP-N-acetylglucosamine + citronellyl-lipid I

UDP + N-acetylglucosamine-citronellyl-lipid I

UDP-N-acetylglucosamine + lipid I

UDP + lipid II

-

i.e. Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

658448, 659026

i.e. N-acetylglucosamine-(1-4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(6E,10E,14Z)-2,6,10,14-tetramethyl-hexadeca-2,6,10,14-tetraene

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(6E,10E,14Z)-2,6,10,14-tetramethyl-hexadeca-2,6,10,14-tetraene

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(7S)-2,7-dimethyloct-2-ene

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(7S)-2,7-dimethyloct-2-ene

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-p-nitrophenol

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-p-nitrophenol

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphobutane

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphobutane

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphododecane

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphododecane

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoicosane

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoicosane

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphotetradecane

UDP + GlcNAcbeta(1-4)Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphotetradecane

-

-

?

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + GlcNAc-(1-4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + N-acetylglucosamine-(1-4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

4 entries

UDP-N-acetylglucosamine + MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydrodecaprenol

UDP + N-acetylglucosamine-MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydrodecaprenol

-

very poor substrate

-

?

UDP-N-acetylglucosamine + MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydroheptaprenol

UDP + N-acetylglucosamine-MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydroheptaprenol

-

-

r

UDP-N-acetylglucosamine + MurNAc(Neta-dansylpentapeptide)-diphosphoryl (R,S-alpha-dihydroheptaprenol)

?

-

-

UDP-N-acetylglucosamine + synthetic substrate analogue 2

?

-

synthetic substrate analogue 2: a 10 carbon citronellyl derivative

-

?

UDP-N-acetylglucosamine + synthetic substrate analogue 7

?

-

synthetic substrate analogue 7: a derivative containing a 20 carbon chain with a cis-allylic double bond

-

?

additional information

?

-

12 entries

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + GlcNAc-(1-4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP-N-acetylglucosamine + Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

UDP + N-acetylglucosamine-(1-4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

additional information

?

-

-

role of enzyme in peptidoglycan biosynthesis pathway

659026

-

Mg2+

-

activates

Mn2+

-

activating compound

(1R,3R,3aS,6aR)-1-{[2-({[(2R,3R,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl] methyl}amino)-2-oxoethyl]carbamoyl}-3-(2-methoxyphenyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrol-2-ium

-

(1R,3S,3aR,6aS)-3-((S)-1,2-dihydroxyethyl)-N-(2-((((2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-2-oxoethyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrole-1-carboxamide

-

(1R,3S,3aR,6aS)-3-(2,3-dimethoxyphenyl)-N-[2-({[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}amino)-2-oxoethyl]-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrole-1-carboxamide

-

(1R,3S,3aR,6aS)-N-[2-({[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}amino)-2-oxoethyl]-1-methyl-4,6-dioxo-3,5-diphenyloctahydropyrrolo[3,4-c]pyrrole-1-carboxamide

-

(1R,3S,3aR,6aS)-N-[2-({[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}amino)-2-oxoethyl]-3-(2-methoxyphenyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrole-1-carboxamide

-

(1R,3S,3aR,6aS)-N-[2-({[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}amino)-2-oxoethyl]-3-(2-methoxyphenyl)-2-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrole-1-carboxamide

-

(2S)-N-{[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}-1-{[(1R,3S,3aR,6aS)-3-(2-methoxyphenyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrol-1-yl]carbonyl}pyrrolidine-2-carboxamide

-

(2S)-N-{[(2R,3S,4R,5R)-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-3,4-dihydroxytetrahydrofuran-2-yl]methyl}-1-{[(1R,3S,3aR,6aS)-5-ethyl-3-(2-methoxyphenyl)-1-methyl-4,6-dioxooctahydropyrrolo[3,4-c]pyrrol-1-yl]carbonyl}pyrrolidine-2-carboxamide

-

(4R,4aR,5aS,6R,12aS)-4-(dimethylamino)-1,5,10,12,12a-pentahydroxy-6-methyl-3,11-dioxo-3,4,4a,5,5a,6,11,12a- octahydrotetracene-2-carboxamide

-

({2-[3-(4-methoxybenzyl)-6-(methoxycarbonyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl]-2-oxoethyl}sulfanyl)acetic acid

-

2-(carboxyethynyl)-3,4-dihydroxybenzoic acid

-

2-(phosphonooxy)butanoic acid

-

4-{[(1S,11aS)-5,11-dioxo-7-phenyl-2,3,5,10,11,11a-hexahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-1-yl]amino}-4-oxobutanoic acid

-

5'-({[(2R,3R,3aR,6aR)-3-carboxy-2-(2,3-dihydroxypropyl)-6a-methyl-4,6-dioxohexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-({[(2S,3R,3aR,6aR)-3-carboxy-2-(2-methoxyphenyl)-6a-methyl-4,6-dioxohexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-({[(2S,3R,3aS,6aR)-3-carboxy-2-(2,3-dimethoxyphenyl)-6a-methyl-4,6-dioxohexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-({[(2S,3R,3aS,6aR)-3-carboxy-2-(2-methoxyphenyl)-6a-methyl-4,6-dioxohexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-({[(2S,3R,3aS,6aR)-3-carboxy-2-(3,4-dimethoxyphenyl)-6a-methyl-4,6-dioxohexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-({[(2S,3R,3aS,6aR)-3-carboxy-6a-methyl-4,6-dioxo-2-phenylhexahydropyrrolo[3,4-b]pyrrol-5(1H)-yl]acetyl}amino)-5'-deoxyuridine

-

5'-deoxy-5'-[(N-{[(2R,3R,4R,5R)-3-(methoxycarbonyl)-5-(2-methoxyphenyl)-2,4-dimethylpyrrolidinium-2-yl]carbonyl}glycyl) amino]uridine

-

5'-O-(3-{[(1R,3S,3aR,6aS)-3-(2-methoxyphenyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrol-1-yl]methoxy}-3-oxopropanoyl)uridine

-

5'-O-(4-{[(1R,3S,3aR,6aS)-3-(2-methoxyphenyl)-1-methyl-4,6-dioxo-5-phenyloctahydropyrrolo[3,4-c]pyrrol-1-yl]methoxy}-4-oxobutanoyl)uridine

-

5'-[(1-{[(2R,3R,4R,5S)-3,4-bis(methoxycarbonyl)-5-(2-methoxyphenyl)-2-methylpyrrolidin-2-yl]carbonyl}-L-prolyl)amino]-5'-deoxyuridine

-

5'-[(1-{[(2R,3S,4R,5S)-3,4-bis(methoxycarbonyl)-5-(2-methoxyphenyl)-2-methylpyrrolidin-2-yl]carbonyl}-L-prolyl)amino]-5'-deoxyuridine

-

5'-{[(3S,4R,5S)-3,4-bis(methoxycarbonyl)-5-(2-methoxyphenyl)-2-methyl-D-prolylglycyl]amino}-5'-deoxyuridine

-

5'-{[(3S5'-{[(3S,4R,5S)-3,4-bis(methoxycarbonyl)-2-methyl-5-phenyl-D-prolylglycyl]amino}-5'-deoxyuridine,4R,5S)-3,4-bis(methoxycarbonyl)-2-methyl-5-phenyl-D-prolylglycyl]amino}-5'-deoxyuridine

-

cephalosporin C

-

50% inhibition at 0.014 mg/ml

EDTA

-

methanol

-

Moenomycin

-

50% inhibition at 0.0106 mM

murgocil

Staphylococcus aureus

-

a highly bioactive staphylococcal-specific inhibitor of intracellular membrane-associated enzyme MurG

735347

N-({6-[(4-cyano-2-fluorobenzyl)oxy]naphthalen-2-yl}sulfonyl)-D-glutamic acid

-

N-p-chlorobiphenyl-vancomycin

-

50% inhibition at 0.00125 mM

NG-p-chlorobiphenyl-vancomycin

-

50% inhibition at 0.0214 mM

Ni2+

-

nisin

-

50% inhibition at 0.016 mg/ml

ramoplanin

trifluoroethanol

-

tunicamycin

UDP

Vancomycin

Zn2+

-

[1-hydroxy-2-(pyridin-3-yl)ethane-1,1-diyl]bis(phosphonic acid)

-

additional information

4 entries

-

Ca2+

-

cardiolipin

-

the activity of MurG is increased in the presence of cardiolipin

dimethyl sulfoxide

-

DMSO

-

high concentration (35%) of dimethylsulfoxide is necessary for maximal enzyme activity

Mg2+

-

488971, 488973, 488974, 488976, 488978

1.5

2'-dUDP-N-acetyl-alpha-D-glucosamine

-

488977

0.0028 - 0.037

biotin-labelled lipid I analogue

-

0.036 - 0.044

biotinylated citronellyl-lipid I

0.053

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(6E,10E,14Z)-2,6,10,14-tetramethyl-hexadeca-2,6,10,14-tetraene

-

pH 7.9, 37°C

0.047

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-(7S)-2,7-dimethyloct-2-ene

-

pH 7.9, 37°C

0.024

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-p-nitrophenol

-

pH 7.9, 37°C

0.022

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphobutane

-

pH 7.9, 37°C

0.029

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphododecane

-

pH 7.9, 37°C

0.052

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoicosane

-

pH 7.9, 37°C

0.039

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphotetradecane

-

pH 7.9, 37°C

0.02

Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphosphoundecaprenol

-

pH 7.9, 37°C

0.0028

MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydroheptaprenol

-

0.0028 - 0.011

MurNAc(Neta-dansylpentapeptide)-diphosphoryl (R,S-alpha-dihydroheptaprenol)

9 entries

0.553

synthetic substrate analogue 2

-

10 carbon citronellyl derivative

-

0.053

synthetic substrate analogue 7

-

0.0098 - 1.03

UDP-GlcNAc

10 entries

0.045 - 0.15

UDP-N-acetylglucosamine

6 entries

additional information

-

continous fluorescence coupled enzyme assay method

-

0.123 - 0.317

biotinylated citronellyl-lipid I

0.933

MurNAc(Nepsilon-dansylpentapeptide)-pyrophosphoryl (R,S)-alpha-dihydroheptaprenol

-

2.23

synthetic substrate analogue 2

-

10 carbon citronellyl derivative

-

14

synthetic substrate analogue 7

-

0.001 - 0.72

UDP-GlcNAc

8 entries

0.27 - 0.93

UDP-N-acetylglucosamine

0.000000096

-

thermosensitive murG mutant strain GS58, 40°C

0.000000336

-

thermosensitive murG mutant strain GS58, 30°C

0.0000016

-

parental strain, 30°C

0.000002

-

parental strain, 40°C

0.0000064

-

murG gene under the control of the lac promotor

0.0000141

-

0.000019

-

mutant N127A

0.000063

-

mutant E268A

0.00026

-

mutant H18A

0.00028

-

mutant Q288A

0.00043

-

mutant E125A

0.00053

-

mutant T15A

0.00067

-

mutant H124A

0.0023

-

mutant R260A

0.0025

-

mutant S191A

0.0064

-

mutant N291A

0.0135

-

mutant N198A

0.071

-

mutant Y105A

0.37

-

wild type enzyme

additional information

7.5

8.3

-

Hepes buffer

7.5 - 9.5

-

22

-

assay at room temperature

25

35

37

-

-

-

Clostridioides difficile R20291

-

-

-

Escherichia coli OV58-pUG18

7 entries

-

488972

-

Micrococcus flavus

gene murG

736947

-

Pseudomonas aeruginosa

-

723716

Q9HW01

UniProt

Staphylococcus aureus

methicillin-resistant

-

-

during exponential growth, MurG localizes to septa of Bacillus subtilis, a zone of active peptidoglycan synthesis, and its N-terminal amphipathic helix is dispensable

-

-

additional information

-

MurG becomes polarly localized when expressed at high cellular concentrations, only at levels that saturate MurGs cellular requirement for growth, the polar MurG is not active

-

722525

malfunction

-

point mutations in a MurG helical causes severe sporulation defects, but does not affect localization nor cause detectable defects during exponential growth. In strains in which the cardiolipin-synthesizing genes are deleted, MurG levels are diminished at the forespore, but MurG localization during sporulation is rescued by external addition of purified cardiolipin. During sporulation, lack of MurG localization heavily affects engulfment dynamics and sporulation efficiency, indicating a defect in MurG enzymatic activity linked to its diffuse localization

metabolism

physiological function

5 entries

additional information

5 entries

Escherichia coli (strain K12)

3s2u, download, 3D-view

SCOPe (3s2u)

CATH (3s2u)

Q9HW01

Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)

37640

-

calculated from the nucleotide sequence, gel filtration

37800

38000

43000

-

determined by SDS-PAGE

76000

-

gel filtration

?

-

x * 38000, SDS-PAGE

658448, 659026

dimer

-

2 * 38000, SDS-PAGE

additional information

hanging-drop vapor-diffusion method, X-ray structure

-

488977

complex of MurG with UDP-GlcNAc, X-ray diffraction structure determination and analysis

Pseudomonas aeruginosa

Q9HW01

723716

F77E

-

site-directed mutagenesis, the mutant shows unaltered localization and cell growth comared to wild-type. Lipid II synthesis might be totally abrogated in mutant F77E explaining why the mutant is blocked in septation, cell wall synthesis is necessary for engulfment

S67E

-

site-directed mutagenesis, the mutant shows unaltered localization and cell growth comared to wild-type

V74E

-

site-directed mutagenesis, the mutant shows unaltered localization and cell growth comared to wild-type. Lipid II synthesis might be totally abrogated in mutant V74E explaining why the mutant is blocked in septation, cell wall synthesis is necessary for engulfment

V81E

-

site-directed mutagenesis, the mutant shows unaltered localization and cell growth comared to wild-type

E125A

-

amino acid substitution for evaluation of its role in the active site of MurG

E268A

H124A

-

amino acid substitution for evaluation of its role in the active site of MurG

H18A

-

amino acid substitution for evaluation of its role in the active site of MurG

L79E/F82E

-

site-directed mutagenesis, the mutant does not bind anionic phospholipids but can localize to the cell poles

722525

N127A

-

amino acid substitution for evaluation of its role in the active site of MurG

N134A

-

amino acid substitution for evaluation of its role in the active site of MurG

N198A

-

amino acid substitution for evaluation of its role in the active site of MurG

N291A

-

amino acid substitution for evaluation of its role in the active site of MurG

Q288A

-

amino acid substitution for evaluation of its role in the active site of MurG

R260A

-

amino acid substitution for evaluation of its role in the active site of MurG

S191A

-

amino acid substitution for evaluation of its role in the active site of MurG

T15A

-

amino acid substitution for evaluation of its role in the active site of MurG

Y105A

-

amino acid substitution for evaluation of its role in the active site of MurG

additional information

-

none of the point mutations shows lethality and cells expressing MurG-GFP point mutants grow normally in absence of the wild-type murG allele

additon of 20% glycerol required for storage at -20°C

-

-70°C

-

-80°C

-

4°C, stable for at least one month

-

-

-

native MurG is extracted from inner membrane vesicles of Escherichia coli cells, His-tagged MurG is purified by Ni-affinity chromatography using His-bind resin

-

676084

Ni2+ affinity column chromatography

-

overexpression in Escherichia coli with His-tag

-

recombinant His6-tagged enzyme from Escherichia coli strain C43 by nickel affinity chromatography

-

using Ni2+-nitrilotriacetate-agarose

-

when overexpressed, enzyme copurifies with lipid vesicles deriving from intracellular vesicular mebranes

-

659026

expressed in Escherichia coli cells BL21-DE3-pLysS

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488971, 488973, 488974, 488976, 488977

gene murG, expression as His6-tagged enzyme in Escherichia coli strain C43

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gene murG, expression of C-terminal YFP-tagged enzyme in a murG-deficient strain of Bacillus subtilis

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into the vector pQE60 for expression of MurG in Escherichia coli JM109 cells

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MurG can be displayed in its active form on phage. Cloned from a pET-21a expression vector (Novagen) into the pFAB5cHis.TT.HUI phagemid vector as a fusion to the 3’-end of a pelB leader sequence and the 5’-end of a truncated gIII