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Information on EC 6.3.5.13 - lipid II isoglutaminyl synthase (glutamine-hydrolysing) for references in articles please use BRENDA:EC6.3.5.13
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EC Tree
IUBMB Comments The enzyme complex, found in Gram-positive bacteria, consists of two subunits. A glutaminase subunit (cf. EC 3.5.1.2, glutaminase) produces an ammonia molecule that is channeled to a ligase subunit, which adds it to the activated D-glutamate residue of lipid II, converting it to an isoglutamine residue.
The enzyme appears in viruses and cellular organisms
Synonyms
Cys-type anaerobic sulfatase-maturating enzyme, GatD, GatD/MurT, MurT/GatD complex, SA1707, SA1708,
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Cys-type anaerobic sulfatase-maturating enzyme
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MurT/GatD complex
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GatD
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GatD/MurT
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SA1707
subunit Gatd
SA1708
subunit Murt
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O = ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
overall reaction
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol = ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
(1b)
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3 = beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
(1c)
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L-glutamine + H2O = L-glutamate + NH3
(1a)
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol:L-glutamine amidoligase (ADP-forming)
The enzyme complex, found in Gram-positive bacteria, consists of two subunits. A glutaminase subunit (cf. EC 3.5.1.2, glutaminase) produces an ammonia molecule that is channeled to a ligase subunit, which adds it to the activated D-glutamate residue of lipid II, converting it to an isoglutamine residue.
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
ATP + glutamate in lipid I + L-glutamine + H2O
ADP + phosphate + glutamine in lipid I + L-glutamate
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?
ATP + glutamate in lipid II + L-glutamine + H2O
ADP + phosphate + glutamine in lipid II + L-glutamate
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ATP + glutamate in lipid II-Gly5 + L-glutamine + H2O
ADP + phosphate + glutamine in lipid II-Gly5 + L-glutamate
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
L-glutamine + H2O
L-glutamate + NH3
additional information
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
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L-glutamine + H2O
L-glutamate + NH3
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L-glutamine + H2O
L-glutamate + NH3
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additional information
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both MurT and GatD are required for the reaction
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additional information
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both MurT and GatD are required for the reaction
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
L-glutamine + H2O
L-glutamate + NH3
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
A0A0H3JN63 and A0A0H3JUU7
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
ADP + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol
A0A0H3JN63 and A0A0H3JUU7
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
A0A0H3JN63 and A0A0H3JUU7
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + ?-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
A0A0H3JN63 and A0A0H3JUU7
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?
ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
Q8DNZ9 and Q8DNZ8
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ATP + beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + L-glutamine + H2O
ADP + phosphate + beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + L-glutamate
Q8DNZ9 and Q8DNZ8
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
A0A0H3JN63 and A0A0H3JUU7
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beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-O-P-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol + NH3
beta-D-GlcNAc-(1->4)-MurNAc-L-Ala-D-isoglutaminyl-L-Lys-D-Ala-D-Ala-diphospho-ditrans,octacis-undecaprenol + phosphate
A0A0H3JN63 and A0A0H3JUU7
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L-glutamine + H2O
L-glutamate + NH3
A0A0H3JN63 and A0A0H3JUU7
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?
L-glutamine + H2O
L-glutamate + NH3
A0A0H3JN63 and A0A0H3JUU7
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0.22
beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
pH 7.5, 30°C
0.078
L-glutamine
pH 7.5, 30°C
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4
beta-D-GlcNAc-(1->4)-Mur2Ac(oyl-L-Ala-gamma-D-Glu-L-Lys-D-Ala-D-Ala)-diphospho-ditrans,octacis-undecaprenol
pH 7.5, 30°C
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A0A0H3JN63 i.e subunit GatD, A0A0H3JUU7 i.e. subunit MurT
UniProt
brenda
Q8DNZ9 i.e. subunit MurT, Q8DNZ8 i.e. subunit GatD
UniProt
brenda
Q8DNZ9 i.e. subunit MurT, Q8DNZ8 i.e. subunit GatD
UniProt
brenda
A0A0H2WZ38 i.e. subunit GatD, A0A0H2WZQ7 i.e. subunit MurT
UniProt
brenda
A0A0H3JN63 i.e subunit GatD, A0A0H3JUU7 i.e. subunit MurT
UniProt
brenda
subunit GatD
UniProt
brenda
A0A0H2WZ38 i.e. subunit GatD, A0A0H2WZQ7 i.e. subunit MurT
UniProt
brenda
subunit GatD
UniProt
brenda
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physiological function
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enzyme complex catalyzes the amidation of the peptidoglycan complex, which is important for polymerization of cell wall building blocks. Inactivation of the GatD active site abolishes lipid II amidation
physiological function
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enzyme complex catalyzes the amidation of the peptidoglycan complex, which is important for polymerization of cell wall building blocks. Inactivation of the GatD active site abolishes lipid II amidation
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MURT_STAAC
Staphylococcus aureus (strain COL)
437
0
49206
Swiss-Prot
MURT_STAAN
437
0
49192
Swiss-Prot
MURT_STRR6
447
0
49576
Swiss-Prot
GATD_STAAC
Staphylococcus aureus (strain COL)
243
0
27430
Swiss-Prot
GATD_STAAN
243
0
27430
Swiss-Prot
GATD_STRR6
260
0
29193
Swiss-Prot
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dimer
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1 * 78800, binary complex MurT/GatD, calculated from sequence
dimer
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1 * 78800, binary complex MurT/GatD, calculated from sequence
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crystal structure of the GatD/MurT complex reveals an open, boomerang-shaped conformation in which GatD is docked onto one end of MurT. Putative active site residues cluster at the interface between GatD and MurT and are contributed by both proteins
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native and the selenomethionine-derivative proteins, crystals diffract beyond 1.85 and 2.25 A , respectively, and belong to space group P212121
structure of subunit GatD, at 1.85 A resolution. A glutamine molecule is found close to the active site funnel, hydrogen-bonded to the conserved residue R128
structure of the MurT/GatD complex at 3.0 A resolution. MurT has central and C-terminal domains similar to Mur ligases with a cysteine-rich insertion, which probably binds zinc, contributing to the interface with GatD. GatD is a glutaminase providing ammonia that is likely channeled to the MurT active site through a cavity network
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C94A
subunit GatD mutant, totally inactive for glutamine deamidation
C94G
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mutation in subunit GaTD, abolishes lipid II amidation
C94S
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subunit GatD mutant, complete loss of activity
D349N
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subunit Murt mutant, displays severely reduced in vitro activity
H189A
subunit GatD mutant, totally inactive for glutamine deamidation
R128A
subunit GatD mutant, totally inactive for glutamine deamidation
C94A
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subunit GatD mutant, totally inactive for glutamine deamidation
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H189A
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subunit GatD mutant, totally inactive for glutamine deamidation
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R128A
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subunit GatD mutant, totally inactive for glutamine deamidation
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C94G
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mutation in subunit GaTD, abolishes lipid II amidation
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C94S
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subunit GatD mutant, complete loss of activity
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D349N
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subunit Murt mutant, displays severely reduced in vitro activity
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recombinant MurT/GatD complex
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expression in Escherichia coli
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Vieira, D.; Figueiredo, T.; Verma, A.; Sobral, R.; Ludovice, A.; De Lencastre, H.; Trincao, J.
Purification, crystallization and preliminary X-ray diffraction analysis of GatD, a glutamine amidotransferase-like protein from Staphylococcus aureus peptidoglycan
Acta Crystallogr. Sect. F
70
632-635
2014
Staphylococcus aureus (A0A0H2WZ38), Staphylococcus aureus COL (A0A0H2WZ38)
brenda
Morlot, C.; Straume, D.; Peters, K.; Hegnar, O.; Simon, N.; Villard, A.; Contreras-Martel, C.; Leisico, F.; Breukink, E.; Gravier-Pelletier, C.; Le Corre, L.; Vollmer, W.; Pietrancosta, N.; Havarstein, L.; Zapun, A.
Structure of the essential peptidoglycan amidotransferase MurT/GatD complex from Streptococcus pneumoniae
Nat. Commun.
9
3180
2018
Streptococcus pneumoniae (Q8DNZ9 and Q8DNZ8), Streptococcus pneumoniae, Streptococcus pneumoniae ATCC BAA-255 (Q8DNZ9 and Q8DNZ8)
brenda
Muench, D.; Roemer, T.; Lee, S.H.; Engeser, M.; Sahl, H.G.; Schneider, T.
Identification and in vitro analysis of the GatD/MurT enzyme-complex catalyzing lipid II amidation in Staphylococcus aureus
PLoS Pathog.
8
e1002509
2012
Staphylococcus aureus (A0A0H3JN63 and A0A0H3JUU7), Staphylococcus aureus, Staphylococcus aureus N315 (A0A0H3JN63 and A0A0H3JUU7)
brenda
Noeldeke, E.R.; Muckenfuss, L.M.; Niemann, V.; Mueller, A.; Stoerk, E.; Zocher, G.; Schneider, T.; Stehle, T.
Structural basis of cell wall peptidoglycan amidation by the GatD/MurT complex of Staphylococcus aureus
Sci. Rep.
8
12953
2018
Staphylococcus aureus (A0A0H3JN63 and A0A0H3JUU7), Staphylococcus aureus, Staphylococcus aureus N315 (A0A0H3JN63 and A0A0H3JUU7)
brenda
Leisico, F.; V Vieira, D.; Figueiredo, T.A.; Silva, M.; Cabrita, E.J.; Sobral, R.G.; Ludovice, A.M.; Trincao, J.; Romao, M.J.; de Lencastre, H.; Santos-Silva, T.
First insights of peptidoglycan amidation in Gram-positive bacteria - the high-resolution crystal structure of Staphylococcus aureus glutamine amidotransferase GatD
Sci. Rep.
8
5313
2018
Staphylococcus aureus (A0A0H2WZ38 and A0A0H2WZQ7), Staphylococcus aureus COL (A0A0H2WZ38 and A0A0H2WZQ7)
brenda
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6.3.5.13 lipid II isoglutaminyl synthase (glutamine-hydrolysing)
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