Cloned (Comment) | Organism |
---|---|
gene kamA, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS | Clostridium subterminale |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | [4Fe-4S] cluster | Clostridium subterminale |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine | Clostridium subterminale | - |
(3S)-3,6-diaminohexanoate | - |
? | |
L-lysine | Clostridium subterminale SB4 | - |
(3S)-3,6-diaminohexanoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridium subterminale | Q9XBQ8 | - |
- |
Clostridium subterminale SB4 | Q9XBQ8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography, ultrafiltration, and dialysis | Clostridium subterminale |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
L-lysine = (3S)-3,6-diaminohexanoate | lysine 2,3-aminomutase (LAM) is a radical S-adenosyl-L-methionine (SAM) enzyme, catalysis is initiated by reductive cleavage of the S-adenosyl-L-methionine S-C5' bond, which creates the highly reactive 5'-deoxyadenosyl radical, the same radical generated by homolytic Co-C bond cleavage in B12 radical enzymes. The S-adenosyl-L-methionine surrogate S-3',4'-anhydroadenosyl-L-methionine can replace S-adenosyl-L-methionine as a cofactor in the isomerization of L-alpha-lysine to L-beta-lysine by 2,3-LAM, via the stable allylic anhydroadenosyl radical. The holoenzyme coordinates a pyridoxal 5'-phosphate cofactor through formation of an internal aldimine with Lys337. As L-alpha-lysine binds, pyridoxal 5'-phosphate forms an external aldimine linkage to the alpha-amine group of the substrate. Reductive cleavage of S-adenosyl-L-methionine leads to formation of 5'-dA radical. Electron transfer from the [4Fe4S]1+ cluster initiates radical S-adenosyl-L-methionine reactions by reductive cleavage of the S-C5' bond to create the highly reactive 5'-deoxyadenosyl radical | Clostridium subterminale |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-lysine | - |
Clostridium subterminale | (3S)-3,6-diaminohexanoate | - |
? | |
L-lysine | - |
Clostridium subterminale SB4 | (3S)-3,6-diaminohexanoate | - |
? | |
additional information | substrate 13C ENDOR measurements | Clostridium subterminale | ? | - |
? | |
additional information | substrate 13C ENDOR measurements | Clostridium subterminale SB4 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
kamA | - |
Clostridium subterminale |
LAM | - |
Clostridium subterminale |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Clostridium subterminale |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Clostridium subterminale |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | S-3',4'-anhydroadenosyl-L-methionine, anSAM, development of the S-adenosyl-L-methionine surrogate, the compound is a true cofactor for lysine 2,3-aminomutase. Specific activity is 0.1 IU/mg | Clostridium subterminale | |
pyridoxal 5'-phosphate | - |
Clostridium subterminale | |
S-adenosyl-L-methionine | SAM, natural cofactor, lysine 2,3-aminomutase (LAM) is a radical S-adenosyl-L-methionine (SAM) enzyme. Enzyme 2,3-LAM utilizes radical-generating machinery comprising SAM anchored to the unique Fe2+ of a [4Fe-4S] cluster via a classical five-membered N,O chelate ring. Specific activity is 35-40 IU/mg | Clostridium subterminale | |
[4Fe-4S] cluster | - |
Clostridium subterminale |
General Information | Comment | Organism |
---|---|---|
evolution | lysine 2,3-aminomutase (LAM) is a member of the radical S-adenosyl-L-methionine (SAM) enzyme superfamily whose reactions are initiated by radical-generating machinery comprising SAM anchored to the unique Fe of a [4Fe-4S] cluster via a classical five-membered N,O chelate ring formed by the methionine | Clostridium subterminale |
additional information | substitution of SAM with S-3',4'-anhydroadenosyl-L-methionine leads to generation of a stable allylic analogue of 5'-dA. radical. Deuterium labeling at positions 2', 3', and 5' dramatically alters the continuous-wave (CW) EPR spectrum | Clostridium subterminale |
physiological function | lysine 2,3-aminomutase (LAM) utilizes the radical-SAM machinery to isomerize L-alpha-lysine to L-beta-lysine | Clostridium subterminale |