Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Salmonella enterica subsp. enterica serovar Typhimurium |
Protein Variants | Comment | Organism |
---|---|---|
D45N | below 0.02% of the amidation activity of wild-type, no change in the order of amidation | Salmonella enterica subsp. enterica serovar Typhimurium |
D48N | 0.2% of the amidation activity of wild-type, no change in the order of amidation | Salmonella enterica subsp. enterica serovar Typhimurium |
D97N | 32.4% of the amidation activity of wild-type, no change in the order of amidation | Salmonella enterica subsp. enterica serovar Typhimurium |
E90Q | below 0.02% of the amidation activity of wild-type, no change in the order of amidation | Salmonella enterica subsp. enterica serovar Typhimurium |
L47A | 10.6% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate | Salmonella enterica subsp. enterica serovar Typhimurium |
Y46A | 3.8% of the amidation activity of wild-type, specific decrease in the affinity of the enzyme for the c-monoamide intermediate | Salmonella enterica subsp. enterica serovar Typhimurium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00074 | - |
cobyrinate | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
0.0027 | - |
ATP | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
0.053 | - |
L-glutamine | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
26.2 | - |
ammonia | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
50000 | - |
1 * 50039, calculated, x * 50000, SDS-PAGE | Salmonella enterica subsp. enterica serovar Typhimurium |
50039 | - |
1 * 50039, calculated, x * 50000, SDS-PAGE | Salmonella enterica subsp. enterica serovar Typhimurium |
53000 | - |
gel filtration | Salmonella enterica subsp. enterica serovar Typhimurium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Salmonella enterica subsp. enterica serovar Typhimurium | - |
LT2 | - |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O = 2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate | catalytic mechanism in which CbiA catalyzes the amidation of the c-carboxylate, and then the intermediate is released into solution and binds to the same catalytic site for the amidation of the a-carboxylate | Salmonella enterica subsp. enterica serovar Typhimurium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 ATP + cobyrinate + 2 ammonia + 2 H2O | - |
Salmonella enterica subsp. enterica serovar Typhimurium | 2 ADP + 2 phosphate + cobyrinate a,c-diamide | - |
? | |
2 ATP + cobyrinate + 2 L-glutamine + 2 H2O | - |
Salmonella enterica subsp. enterica serovar Typhimurium | 2 ADP + 2 phosphate + cobyrinate a,c-diamide + 2 L-glutamate | the hydrolysis of glutamine and the synthesis of the cobyrinic acid a,c-diamide product are uncoupled. CbiA catalyzes the sequential amidation of the c- and a-carboxylate groups of cobyrinic acid via the formation of a phosphorylated intermediate. CbiA catalyzes the amidation of the c-carboxylate, and then the intermediate is released into solution and binds to the same catalytic site for the amidation of the a-carboxylate | ? | |
additional information | CbiA is able to hydrolyze glutamine in the absence of the other two substrates. The addition of ATP alone decreases the Km for glutamine by a factor of 2. Cobyrinic acid enhances the kcat for the hydrolysis of glutamine about 2fold and decreases the Km for glutamine by 20fold. The decrease in the Km for glutamine is about 200fold when both ATP and cobyrinic acid are present in the assay solution. There is no change in the Km for glutamine when ADP is added with cobyrinic acid | Salmonella enterica subsp. enterica serovar Typhimurium | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 50039, calculated, x * 50000, SDS-PAGE | Salmonella enterica subsp. enterica serovar Typhimurium |
Synonyms | Comment | Organism |
---|---|---|
CbiA | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.13 | - |
ATP | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
0.16 | - |
L-glutamine | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
0.16 | - |
cobyrinate | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
0.18 | - |
ammonia | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | 8 | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.04 | - |
L-glutamine | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
6.9 | - |
ammonia | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
48.2 | - |
ATP | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium | |
216 | - |
cobyrinate | pH 7.7, 30°C | Salmonella enterica subsp. enterica serovar Typhimurium |