Cloned (Comment) | Organism |
---|---|
gene SlCGT, functional expression in Nicotiana benthamiana leaves using Agrobacterium tumefaciens strain GV2260 transfection method | Solanum lycopersicum |
Protein Variants | Comment | Organism |
---|---|---|
D328A | site-directed mutagenesis, the mutant shows 40% reduced activity | Solanum lycopersicum |
S27A | site-directed mutagenesis, the mutant shows 13% reduced activity | Solanum lycopersicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
PMSF | treatment of purified SlCGT with 1, 10, and 50 mM PMSF leads to strong decreases in caffeoyl transfer activities to 60%, 40%, and 0% activity, respectively | Solanum lycopersicum |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
apoplast | apoplastic space of tomato leaves | Solanum lycopersicum | 48046 | - |
additional information | SiCGT contains a putative targeting signal of 24 amino acids indicating an entry of the SlCGT into the secretory pathway | Solanum lycopersicum | - |
- |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
39950 | - |
x * 40000, about, SDS-PAGE, x * 39950, mature CGT without signal sequence, sequence calculation, x * 42580, CGT with signal sequence, sequence calculation | Solanum lycopersicum |
40000 | - |
x * 40000, about, SDS-PAGE, x * 39950, mature CGT without signal sequence, sequence calculation, x * 42580, CGT with signal sequence, sequence calculation | Solanum lycopersicum |
42580 | - |
x * 40000, about, SDS-PAGE, x * 39950, mature CGT without signal sequence, sequence calculation, x * 42580, CGT with signal sequence, sequence calculation | Solanum lycopersicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
chlorogenate + glucarate | Solanum lycopersicum | the enzyme catalyzes the formation of caffeoylglucarate and caffeoylgalactarate using chlorogenate, i.e. 5-O-caffeoylquinate, as acyl donor | quinate + 2-O-caffeoylglucarate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Solanum lycopersicum | E7AIM3 | cv. Moneymaker, gene SlCGT, single-copy gene | - |
Purification (Comment) | Organism |
---|---|
native SlCGT 585fold from seedlings by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, ultrafiltration, gel filtration, and again anion exchange chromatography | Solanum lycopersicum |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
flower | - |
Solanum lycopersicum | - |
fruit | at different stages of ripening | Solanum lycopersicum | - |
leaf | apoplastic space of tomato leaves | Solanum lycopersicum | - |
additional information | tissue expression pattern, overview. Transcripts of SlCGT, enzyme activity, and caffeoylglucarate accumulate preferentially in tomato leaves and flowers | Solanum lycopersicum | - |
root | - |
Solanum lycopersicum | - |
seedling | - |
Solanum lycopersicum | - |
stem | - |
Solanum lycopersicum | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.45 | - |
purified native enzyme, pH 6.0, 30°C | Solanum lycopersicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
chlorogenate + glucarate | - |
Solanum lycopersicum | quinate + 2-O-caffeoylglucarate | - |
? | |
chlorogenate + glucarate | the enzyme catalyzes the formation of caffeoylglucarate and caffeoylgalactarate using chlorogenate, i.e. 5-O-caffeoylquinate, as acyl donor | Solanum lycopersicum | quinate + 2-O-caffeoylglucarate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40000, about, SDS-PAGE, x * 39950, mature CGT without signal sequence, sequence calculation, x * 42580, CGT with signal sequence, sequence calculation | Solanum lycopersicum |
More | peptide sequences are obtained by trypsin digestion and spectrometric sequencing, structure-function analysis, overview | Solanum lycopersicum |
Synonyms | Comment | Organism |
---|---|---|
CGT | - |
Solanum lycopersicum |
chlorogenate:glucarate caffeoyltransferase | - |
Solanum lycopersicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Solanum lycopersicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Solanum lycopersicum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Solanum lycopersicum | sequence calculation, mature CGT without signal sequence | - |
5.01 |
Solanum lycopersicum | sequence calculation, CGT with signal sequence | - |
5.25 |
General Information | Comment | Organism |
---|---|---|
evolution | the sequence determines SlCGT as a GDSL lipase-like protein, representing it as a member of the SGNH protein superfamily. Lipases of this family employ a catalytic triad of Ser-Asp-His with Ser as nucleophile of the GDSL motif. The catalytic triad of the GDSL lipase is not essential for SlCGT enzymatic activity. SlCGT is therefore an example of a GDSL lipase-like protein that has lost hydrolytic activity and has acquired a completely new function in plant metabolism, functioning in secondary metabolism as acyltransferase in synthesis of hydroxycinnamate esters by employing amino acid residues different from the lipase catalytic triad | Solanum lycopersicum |
additional information | the deduced SlCGT amino acid sequence matches the conservation of the serine catalytic motif GXSXXDXG within the first consensus block near the N-terminus. A serine moiety of the GDSL protein may be part of the catalytic center in SlCGT | Solanum lycopersicum |