Cloned (Comment) | Organism |
---|---|
sequence comparisons and phylogenetic tree, recombinant expression of His-tagged chimeric enzyme mutants in Escherichia coli | Pisum sativum |
sequence comparisons and phylogenetic tree, recombinant expression of His-tagged chimeric enzyme mutants in Escherichia coli | Arabidopsis thaliana |
Protein Variants | Comment | Organism |
---|---|---|
247H/Ins254A/R259W | mutant is able to methylate both Rubisco and fructose 1,6-bisphosphate aldolase with similar efficiency | Arabidopsis thaliana |
I242V | mutant is not able to methylate Rubisco but rather behaves as wild-type LSMT | Arabidopsis thaliana |
Ins254A | enzyme is able to methylate Rubisco in addition to fructose 1,6-bisphosphate aldolase | Arabidopsis thaliana |
Ins254A/R259W | mutation enhances methylation of Rubisco without altering fructose 1,6-bisphosphate aldolase methylation | Arabidopsis thaliana |
additional information | construction of chimera between Pisum sativum and Arabidopsis thaliana enzymes to localize regions of the enzymes responsible for the observed difference in substrate specificity. Contrary to the Pisum sativum enzyme, Arabidopsis thaliana LSMT is not able to trimethylate Rubisco | Pisum sativum |
additional information | construction of chimera between Pisum sativum and Arabidopsis thaliana enzymes to localize regions of the enzymes responsible for the observed difference in substrate specificity. Contrary to the Pisum sativum enzyme, Arabidopsis thaliana LSMT is not able to trimethylate Rubisco | Arabidopsis thaliana |
additional information | identification of an LSMT region important for RBCL methylation by domain swapping, Arabidopsis thaliana AtLSMT and Pisum sativum PsLSMT are selected as representatives of monofunctional and bifunctional enzymes, respectively. An initial set of chimeric enzymes (chimera 1 and 2) is constructed in which the N-terminal portion of one parental mature enzyme, i.e. devoid of its amino terminal sequence for targeting to plastids, is replaced by the equivalent of the second parental enzyme, and vice versa. The exchanged peptide fragment comprises the nSET domain, the N-terminal part of the SET domain, and the entire iSET domain of both model enzymes. Chimera 1, bearing the N-terminal portion from PsLSMT, displays an AtLSMT-like activity, methylating only FBA2, while its reciprocal counterpart, chimera 2, displays a PsLSMT-like profile, methylating both FBA2 and RBCL. For chimera 2, measured activities are somewhat impaired compared with those of the PsLSMT parent enzyme. Chimera 1 and 2 confirm that the iSET domain is not critical for LSMT substrate selectivity. Phenotypes overview | Pisum sativum |
additional information | identification of an LSMT region important for RBCL methylation by domain swapping, Arabidopsis thaliana AtLSMT and Pisum sativum PsLSMT are selected as representatives of monofunctional and bifunctional enzymes, respectively. An initial set of chimeric enzymes (chimera 1 and 2) is constructed in which the N-terminal portion of one parental mature enzyme, i.e. devoid of its amino terminal sequence for targeting to plastids, is replaced by the equivalent of the second parental enzyme, and vice versa. The exchanged peptide fragment comprises the nSET domain, the N-terminal part of the SET domain, and the entire iSET domain of both model enzymes. Chimera 1, bearing the N-terminal portion from PsLSMT, displays an AtLSMT-like activity, methylating only FBA2, while its reciprocal counterpart, chimera 2, displays a PsLSMT-like profile, methylating both FBA2 and RBCL. For chimera 2, measured activities are somewhat impaired compared with those of the PsLSMT parent enzyme. Chimera 1 and 2 confirm that the iSET domain is not critical for LSMT substrate selectivity. Phenotypes overview | Arabidopsis thaliana |
P240A | mutant is not able to methylate Rubisco but rather behaves as wild-type LSMT | Arabidopsis thaliana |
R259W | mutant is not able to methylate Rubisco but rather behaves as wild-type LSMT | Arabidopsis thaliana |
S299P | mutant is not able to methylate Rubisco but rather behaves as wild-type LSMT | Arabidopsis thaliana |
V269L | mutant is not able to methylate Rubisco but rather behaves as wild-type LSMT | Arabidopsis thaliana |
Y247H | mutant is not able to methylate Rubisco, activity is severely impaired | Arabidopsis thaliana |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Pisum sativum | 9507 | - |
chloroplast | - |
Arabidopsis thaliana | 9507 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
3 adenosyl-L-methionine + [Rubisco large subunit]-L-lysine | Pisum sativum | - |
3 S-adenosyl-L-homocysteine + [Rubisco large subunit]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [fructose-bisphosphate aldolase]-L-lysine | Pisum sativum | - |
3 S-adenosyl-L-homocysteine + [fructose-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [fructose-bisphosphate aldolase]-L-lysine | Arabidopsis thaliana | - |
3 S-adenosyl-L-homocysteine + [fructose-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | Q9XI84 | - |
- |
Pisum sativum | Q43088 | - |
- |
Pisum sativum | Q43088 | cf. EC 2.1.1.127 | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged chimeric enzyme mutants from Escherichia coli by nickel affinity chromatography | Pisum sativum |
recombinant His-tagged chimeric enzyme mutants from Escherichia coli by nickel affinity chromatography | Arabidopsis thaliana |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Pisum sativum | - |
leaf | - |
Arabidopsis thaliana | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
3 adenosyl-L-methionine + [Rubisco large subunit]-L-lysine | - |
Pisum sativum | 3 S-adenosyl-L-homocysteine + [Rubisco large subunit]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 adenosyl-L-methionine + [Rubisco large subunit]-L-lysine | i.e. RBCL | Pisum sativum | 3 S-adenosyl-L-homocysteine + [Rubisco large subunit]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [chloroplastic fructose 1,6-bisphosphate aldolase isoform 2]-L-lysine394 | - |
Arabidopsis thaliana | 3 S-adenosyl-L-homocysteine + [chloroplastic fructose 1,6-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine394 | - |
? | |
3 S-adenosyl-L-methionine + [chloroplastic fructose 1,6-bisphosphate aldolase]-L-lysine | - |
Pisum sativum | 3 S-adenosyl-L-homocysteine + [chloroplastic fructose 1,6-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [fructose-bisphosphate aldolase]-L-lysine | - |
Pisum sativum | 3 S-adenosyl-L-homocysteine + [fructose-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [fructose-bisphosphate aldolase]-L-lysine | - |
Arabidopsis thaliana | 3 S-adenosyl-L-homocysteine + [fructose-bisphosphate aldolase]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
3 S-adenosyl-L-methionine + [ribulose-1,5-bisphosphate-carboxylase]-lysine | - |
Pisum sativum | 3 S-adenosyl-L-homocysteine + [ribulose-1,5-bisphosphate-carboxylase]-N6,N6,N6-trimethyl-L-lysine | - |
? | |
additional information | no trimethylation of the Rubisco large subunit by the enzyme from Arabidosis thaliana | Arabidopsis thaliana | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
chloroplastic protein methyltransferase | - |
Pisum sativum |
chloroplastic protein methyltransferase | - |
Arabidopsis thaliana |
large subunit Rubisco methyltransferase | - |
Pisum sativum |
LSMT | - |
Pisum sativum |
LSMT | - |
Arabidopsis thaliana |
PsLSMT | - |
Pisum sativum |
rbcMT | - |
Pisum sativum |
rbcMT | - |
Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Pisum sativum | |
S-adenosyl-L-methionine | - |
Arabidopsis thaliana |
General Information | Comment | Organism |
---|---|---|
evolution | molecular evolution of the substrate specificity of chloroplastic aldolases/Rubisco lysine methyltransferases in plants, overview. The His-Ala/Pro-Trp triad located in the central part of LSMT enzymes is the key motif to confer the capacity to trimethylate Rubisco. Two of the critical residues are located on a surface loop outside the methyltransferase catalytic site. A strict correlation between the presence of the triad motif and the in vivo methylation status of Rubisco is observed, distribution of the motif into a phylogenetic tree, overview. Chloroplastic fructose-1,6-bisphosphate aldolases (FBAs) are naturally trimethylated in both Pisum sativum and Arabidopsis thaliana, whereas the Rubisco large subunit is trimethylated only in the former species. The distribution of the motif into a phylogenetic tree further suggests that the ancestral function of LSMT was FBA trimethylation. In a recent event during higher plant evolution, this function evolved in ancestors of Fabaceae, Cucurbitaceae, and Rosaceae to include Rubisco as an additional substrate to the archetypal enzyme | Pisum sativum |
evolution | molecular evolution of the substrate specificity of chloroplastic aldolases/Rubisco lysine methyltransferases in plants, overview. The His-Ala/Pro-Trp triad located in the central part of LSMT enzymes is the key motif to confer the capacity to trimethylate Rubisco. Two of the critical residues are located on a surface loop outside the methyltransferase catalytic site. A strict correlation between the presence of the triad motif and the in vivo methylation status of Rubisco is observed, distribution of the motif into a phylogenetic tree, overview. Chloroplastic fructose-1,6-bisphosphate aldolases (FBAs) are naturally trimethylated in both Pisum sativum and Arabidopsis thaliana, whereas the Rubisco large subunit is trimethylated only in the former species. The distribution of the motif into a phylogenetic tree further suggests that the ancestral function of LSMT was FBA trimethylation. In a recent event during higher plant evolution, this function evolved in ancestors of Fabaceae, Cucurbitaceae, and Rosaceae to include Rubisco as an additional substrate to the archetypal enzyme | Arabidopsis thaliana |
additional information | structure of PsLSMT enzyme in complex with the AdoMet structural analogue aza-adenosyl-Lmethionine (PDB ID 2H2E), overview | Pisum sativum |