Literature summary for 6.3.2.4 extracted from

Shi, Y.; Walsh, C.T.
Active site mapping of Escherichia coli D-Ala-D-Ala ligase by structure-based mutagenesis (1995), Biochemistry, 34, 2768-2776.

Protein Variants

Protein Variants	Comment	Organism
additional information	mutant enzymes at K144, K215, and E270 in the ATP binding site, at E15, S150, H63, and R255 in the first D-Ala subsite, and at Y216, S281, L282, and D257 in the second D-Ala subsite. Mutant enzymes E270Q, K215A, R255A, and D257N retain very low or no detectable activity. Mutants enzyme Y216F shows substantial retention of activity	Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
Phosphinates	-	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0012	-	D-Ala	N-terminal Ala, wild-type enzyme	Escherichia coli
0.008	-	ATP	mutant enzyme Y216K	Escherichia coli
0.033	-	ATP	mutant enzyme Y216F	Escherichia coli
0.049	-	ATP	wild-type enzyme	Escherichia coli
0.055	-	ATP	mutant enzyme E15Q	Escherichia coli
0.161	-	ATP	mutant enzyme H63Q	Escherichia coli
0.179	-	ATP	mutant enzyme S150A	Escherichia coli
0.26	-	D-Ala	N-terminal Ala, mutant enzyme Y216F	Escherichia coli
0.43	-	D-Ala	N-terminal Ala, mutant enzyme E15Q, mutant enzyme Y216K	Escherichia coli
0.49	-	D-Ala	N-terminal Ala, mutant enzyme L282R	Escherichia coli
0.55	-	D-Ala	N-terminal Ala, mutant enzyme H63Q	Escherichia coli
0.76	-	D-Ala	N-terminal Ala, mutant enzyme S150A	Escherichia coli
1.05	-	D-Ala	C-terminal Ala, mutant enzyme D257N	Escherichia coli
1.13	-	D-Ala	C-terminal Ala, wild-type enzyme	Escherichia coli
2.4	-	ATP	mutant enzyme K144A	Escherichia coli
2.5	-	ATP	mutant enzyme K144T	Escherichia coli
5.4	-	D-Ala	reaction with D-Met + ATP	Escherichia coli
6	-	D-Phe	reaction with D-Ala + ATP	Escherichia coli
6.4	-	D-Ala	reaction with D-Phe + ATP	Escherichia coli
9	-	D-Met	-	Escherichia coli
15	-	D-norleucine	reaction with D-Ala + ATP	Escherichia coli
20.9	-	D-Ala	C-terminal Ala, mutant enzyme L282R	Escherichia coli
22.8	-	D-Ala	C-terminal Ala, mutant enzyme K144A, reaction with 5 mM ATP	Escherichia coli
22.9	-	D-Ala	C-terminal Ala, mutant enzyme K144A, reaction with 15 mM ATP	Escherichia coli
25.7	-	D-Ala	C-terminal Ala, mutant enzyme Y216F	Escherichia coli
33	-	2-aminopentanoate	reaction with D-Ala + ATP	Escherichia coli
43.7	-	D-Ala	C-terminal Ala, mutant enzyme E15Q	Escherichia coli
57.5	-	D-Ala	C-terminal Ala, mutant enzyme H63Q	Escherichia coli
60.4	-	D-Ala	C-terminal Ala, mutant enzyme S150A	Escherichia coli
70.3	-	D-Ala	C-terminal Ala, mutant enzyme K144T, reaction with 5 mM ATP	Escherichia coli
86	-	D-Ala	C-terminal Ala, mutant enzyme K144T, reaction with 15 mM ATP	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	wild-type and mutant enzyme forms	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + D-Ala + 2-aminopentanoate	-	Escherichia coli	ADP + phosphate + D-Ala-2-aminopentanoate	-	?
ATP + D-Ala + D-Ala	-	Escherichia coli	ADP + phosphate + D-Ala-D-Ala	-	?
ATP + D-Ala + D-Met	-	Escherichia coli	ADP + phosphate + D-Ala-D-Met	-	?
ATP + D-Ala + D-norleucine	-	Escherichia coli	ADP + phosphate + D-Ala-D-norleucine	-	?
ATP + D-Ala + D-Phe	-	Escherichia coli	ADP + phosphate + D-Ala-D-Phe	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	turnover numbers of wild-type and mutant enzyme forms	Escherichia coli