Literature summary for 2.3.1.203 extracted from

Morrison, M.J.; Imperiali, B.
Biochemical analysis and structure determination of bacterial acetyltransferases responsible for the biosynthesis of UDP-N,N-diacetylbacillosamine (2013), J. Biol. Chem., 288, 32248-32260.

Search for more literature for 2.3.1.203

Crystallization (Commentary)

Crystallization (Comment)	Organism
apo-crystal structures of the acetyltransferase domain ATD and in complex with AcCoA. The N-terminal section (Asn199-Leu285) comprises a binding pocket for the UDP-4-amino sugar substrate through a beta-alpha-beta-alpha-beta Rossman fold motif. The C-terminus (Pro286-Leu403) is composed of a left-handed beta-helix motif that, in conjunction with an adjacent PglB-ATD protomer in the trimeric state, forms an extended cleft that is utilized for AcCoA binding	Neisseria gonorrhoeae
compared to isoforms PglD and PglB, WeeI contains an additional loop (Gln174-Pro180) that forms the substrate binding pocket near the pyranose moiety. Residue Gln174 seems to be critical for catalysis. When aligned to the PglD UDP-4-amino structure, this position is analogous to Asn162 that interacts with the carbonyl oxygen of the pyranose C2-acetyl group. In the WeeI apo-structure, Gln174 is within 5 A of the catalytic base His138 and 3.6 A of the AcCoA thioester	Acinetobacter baumannii

Crystallization (Comment)

Organism

apo-crystal structures of the acetyltransferase domain ATD and in complex with AcCoA. The N-terminal section (Asn199-Leu285) comprises a binding pocket for the UDP-4-amino sugar substrate through a beta-alpha-beta-alpha-beta Rossman fold motif. The C-terminus (Pro286-Leu403) is composed of a left-handed beta-helix motif that, in conjunction with an adjacent PglB-ATD protomer in the trimeric state, forms an extended cleft that is utilized for AcCoA binding

Neisseria gonorrhoeae

compared to isoforms PglD and PglB, WeeI contains an additional loop (Gln174-Pro180) that forms the substrate binding pocket near the pyranose moiety. Residue Gln174 seems to be critical for catalysis. When aligned to the PglD UDP-4-amino structure, this position is analogous to Asn162 that interacts with the carbonyl oxygen of the pyranose C2-acetyl group. In the WeeI apo-structure, Gln174 is within 5 A of the catalytic base His138 and 3.6 A of the AcCoA thioester

Acinetobacter baumannii

Protein Variants

Protein Variants	Comment	Organism
D332A	active site mutant, strong loss of activity	Neisseria gonorrhoeae
F13A	active site mutant, strong loss of activity	Acinetobacter baumannii
H210F	active site mutant, strong loss of activity	Neisseria gonorrhoeae
Q174A	active site mutant, strong loss of activity	Acinetobacter baumannii
Q369A	active site mutant, strong loss of activity	Neisseria gonorrhoeae
Q370A	active site mutant, 50% loss of activity	Neisseria gonorrhoeae
T176A	active site mutant, strong loss of activity	Acinetobacter baumannii

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.09	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q370A, pH 7.4, 25°C	Neisseria gonorrhoeae
0.097	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q369A, pH 7.4, 25°C	Neisseria gonorrhoeae
0.099	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Neisseria gonorrhoeae
0.32	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant H210F, pH 7.4, 25°C	Neisseria gonorrhoeae
0.56	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant D332A, pH 7.4, 25°C	Neisseria gonorrhoeae
2.42	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q174A, pH 7.4, 25°C	Acinetobacter baumannii
2.52	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Acinetobacter baumannii
4.51	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant F13A, pH 7.4, 25°C	Acinetobacter baumannii
4.6	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant T176A, pH 7.4, 25°C	Acinetobacter baumannii

Organism

Organism	UniProt	Comment	Textmining
Acinetobacter baumannii	B0V6J7	-	-
Acinetobacter baumannii AYE	B0V6J7	-	-
Neisseria gonorrhoeae	Q5FAE1	-	-
Neisseria gonorrhoeae FA 1090	Q5FAE1	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
acetyl-CoA + UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	-	Neisseria gonorrhoeae	CoA + UDP-2,4-diacetamido-2,4,6-trideoxy-alpha-D-glucopyranose	-	?
acetyl-CoA + UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	-	Acinetobacter baumannii	CoA + UDP-2,4-diacetamido-2,4,6-trideoxy-alpha-D-glucopyranose	-	?
acetyl-CoA + UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	-	Acinetobacter baumannii AYE	CoA + UDP-2,4-diacetamido-2,4,6-trideoxy-alpha-D-glucopyranose	-	?
acetyl-CoA + UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	-	Neisseria gonorrhoeae FA 1090	CoA + UDP-2,4-diacetamido-2,4,6-trideoxy-alpha-D-glucopyranose	-	?

Synonyms

Synonyms	Comment	Organism
ABAYE3807	-	Acinetobacter baumannii
NGO_0085	-	Neisseria gonorrhoeae
WeeI	-	Acinetobacter baumannii

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
1200	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant H210F, pH 7.4, 25°C	Neisseria gonorrhoeae
1900	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q174A, pH 7.4, 25°C	Acinetobacter baumannii
3000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q370A, pH 7.4, 25°C	Neisseria gonorrhoeae
3600	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant D332A, pH 7.4, 25°C	Neisseria gonorrhoeae
5600	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q369A, pH 7.4, 25°C	Neisseria gonorrhoeae
72000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Neisseria gonorrhoeae
82000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant F13A, pH 7.4, 25°C	Acinetobacter baumannii
330000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant T176A, pH 7.4, 25°C	Acinetobacter baumannii
510000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Acinetobacter baumannii

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
800	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q174A, pH 7.4, 25°C	Acinetobacter baumannii
3700	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant H210F, pH 7.4, 25°C	Neisseria gonorrhoeae
6500	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant D332A, pH 7.4, 25°C	Neisseria gonorrhoeae
18000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant F13A, pH 7.4, 25°C	Acinetobacter baumannii
58000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q369A, pH 7.4, 25°C	Neisseria gonorrhoeae
71000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant T176A, pH 7.4, 25°C	Acinetobacter baumannii
200000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Acinetobacter baumannii
340000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	mutant Q370A, pH 7.4, 25°C	Neisseria gonorrhoeae
730000	-	UDP-2-acetamido-4-amino-2,4,6-trideoxy-alpha-D-glucopyranose	wild-type, pH 7.4, 25°C	Neisseria gonorrhoeae