Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | abiotic stess-inducible enzyme, e.g. high salt content | Aphanothece halophytica |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) | Aphanothece halophytica |
orf1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of wild-type and mutant enzymes in Escherichia coli, functional coexpression of the soluble enzyme with the sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, increases the intracellular betaine level and the growth rate of Escherichia coli strain BL21(DE3) | Aphanothece halophytica |
Protein Variants | Comment | Organism |
---|---|---|
R169E | inactive | Aphanothece halophytica |
R169E | site-directed mutagenesis, completely inactive mutant | Aphanothece halophytica |
R169K | site-directed mutagenesis, reduced activity and highly increased Km for ATP with both glycine and sarcosine compared to the wild-type enzyme | Aphanothece halophytica |
R169K | drastic increase of Km of the methyl-acceptor substrates, 2-fold increase in Km for S-adenosyl-L-methionine | Aphanothece halophytica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetate | 80% inhibition at 250 mM, with glycine | Aphanothece halophytica | |
dimethylglycine | versus glycine, 65% inhibition at 250 mM | Aphanothece halophytica | |
ethanolamine | 45% inhibition at 250 mM, with glycine | Aphanothece halophytica | |
KCl | - |
Aphanothece halophytica | |
monomethyl ethanolamine | 35% inhibition at 250 mM, with glycine | Aphanothece halophytica | |
additional information | inhibition rates with sarcosine, overview, no feedback inhibition by betaine up to 2 M, no inhibition with glycine by serine, methionine, asparagine, glutamate, cysteine, and alanine; no inhibition with betaine | Aphanothece halophytica | |
N,N-dimethylglycine | - |
Aphanothece halophytica | |
NaCl | - |
Aphanothece halophytica | |
phenylalanine | 10% inhibition at 250 mM, with glycine | Aphanothece halophytica | |
proline | 30% inhibition at 250 mM, with glycine | Aphanothece halophytica | |
S-adenosyl-L-homocysteine | product inhibition | Aphanothece halophytica | |
sarcosine | competitive inhibition, versus glycine, 70% inhibition at 250 mM | Aphanothece halophytica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.6 | - |
S-adenosyl-L-methionine | pH 8.8, 37°C, recombinant wild-type enzyme, with sarcosine | Aphanothece halophytica | |
0.6 | - |
S-adenosyl-L-methionine | with N-methylglycine as methyl group acceptor, pH 8.8, 37°C | Aphanothece halophytica | |
0.7 | - |
S-adenosyl-L-methionine | pH 8.8, 37°C, recombinant wild-type enzyme, with glycine | Aphanothece halophytica | |
0.7 | - |
S-adenosyl-L-methionine | with glycine as methyl group acceptor, pH 8.8, 37°C | Aphanothece halophytica | |
0.8 | - |
N-methylglycine | pH 8.8, 37°C | Aphanothece halophytica | |
0.8 | - |
sarcosine | pH 8.8, 37°C, recombinant wild-type enzyme | Aphanothece halophytica | |
1 | - |
glycine | pH 8.8, 37°C, recombinant mutant R169K | Aphanothece halophytica | |
1.1 | - |
sarcosine | pH 8.8, 37°C, recombinant mutant R169K | Aphanothece halophytica | |
1.5 | - |
glycine | pH 8.8, 37°C | Aphanothece halophytica | |
1.5 | - |
glycine | pH 8.8, 37°C, recombinant wild-type enzyme | Aphanothece halophytica | |
50 | - |
S-adenosyl-L-methionine | pH 8.8, 37°C, recombinant mutant R169K, with sarcosine | Aphanothece halophytica | |
75 | - |
S-adenosyl-L-methionine | pH 8.8, 37°C, recombinant mutant R169K, with glycine | Aphanothece halophytica |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31210 | - |
calculated from the deduced amino acid sequence | Aphanothece halophytica |
32000 | - |
gel filtration | Aphanothece halophytica |
32000 | - |
recombinant enzyme, gel filtration | Aphanothece halophytica |
33000 | - |
1 * 33000, recombinant enzyme, SDS-PAGE | Aphanothece halophytica |
33000 | - |
1 * 33000, SDS-PAGE, 1 * 32000, gel filtration | Aphanothece halophytica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Aphanothece halophytica | the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress | ? | - |
? | |
S-adenosyl-L-methionine + glycine | Aphanothece halophytica | - |
S-adenosyl-L-homocysteine + sarcosine | - |
? | |
S-adenosyl-L-methionine + glycine | Aphanothece halophytica | synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine | S-adenosyl-L-homocysteine + N-methylglycine | - |
? | |
S-adenosyl-L-methionine + N-methylglycine | Aphanothece halophytica | synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine | S-adenosyl-L-homocysteine + N,N-dimethylglycine | - |
? | |
S-adenosyl-L-methionine + sarcosine | Aphanothece halophytica | - |
S-adenosyl-L-homocysteine + N,N-dimethylglycine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aphanothece halophytica | - |
- |
- |
Aphanothece halophytica | Q83WC4 | inducible enzyme | - |
Purification (Comment) | Organism |
---|---|
- |
Aphanothece halophytica |
recombinant enzyme from Escherichia coli by ammonium sulfate fractionation and ion exchange chromatography to homogeneity | Aphanothece halophytica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress | Aphanothece halophytica | ? | - |
? | |
additional information | enzyme shows strict substrate specificity | Aphanothece halophytica | ? | - |
? | |
S-adenosyl-L-methionine + glycine | - |
Aphanothece halophytica | S-adenosyl-L-homocysteine + sarcosine | - |
? | |
S-adenosyl-L-methionine + glycine | - |
Aphanothece halophytica | S-adenosyl-L-homocysteine + sarcosine | i.e. 2-methylglycine | ? | |
S-adenosyl-L-methionine + glycine | - |
Aphanothece halophytica | S-adenosyl-L-homocysteine + N-methylglycine | - |
? | |
S-adenosyl-L-methionine + glycine | synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine | Aphanothece halophytica | S-adenosyl-L-homocysteine + N-methylglycine | - |
? | |
S-adenosyl-L-methionine + N-methylglycine | - |
Aphanothece halophytica | S-adenosyl-L-homocysteine + N,N-dimethylglycine | - |
? | |
S-adenosyl-L-methionine + N-methylglycine | synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine | Aphanothece halophytica | S-adenosyl-L-homocysteine + N,N-dimethylglycine | - |
? | |
S-adenosyl-L-methionine + sarcosine | - |
Aphanothece halophytica | S-adenosyl-L-homocysteine + N,N-dimethylglycine | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 33000, recombinant enzyme, SDS-PAGE | Aphanothece halophytica |
monomer | 1 * 33000, SDS-PAGE, 1 * 32000, gel filtration | Aphanothece halophytica |
More | enzyme aggregated to tetramers is inactive | Aphanothece halophytica |
Synonyms | Comment | Organism |
---|---|---|
GSMT | - |
Aphanothece halophytica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Aphanothece halophytica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.8 | - |
assay at | Aphanothece halophytica |
9 | - |
glycine or N-methylglycine | Aphanothece halophytica |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
8 | 11 | glycine or N-methylglycine, very low activity below pH 8 | Aphanothece halophytica |