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Literature summary extracted from
- Semisynthesis and initial characterization of sortase A mutants containing selenocysteine and homocysteine (2015), Bioorg. Med. Chem., 23, 2883-2889.
Application
| EC Number | Application | Comment | Organism |
|---|---|---|---|
| 3.4.22.70 | analysis | semisynthetic active site mutant enzymes containing selenocysteine and homocysteine might represent useful tools for further biochemical investigations and engineering approaches of sortases A | Staphylococcus aureus |
| 3.4.22.70 | biotechnology | the bacterial transpeptidase sortase A is a well-established tool in protein chemistry and catalyzes the chemoselective ligation of peptides and proteins | Staphylococcus aureus |
| 3.4.22.70 | synthesis | the bacterial transpeptidase sortase A is a well-established tool in protein chemistry and catalyzes the chemoselective ligation of peptides and proteins | Staphylococcus aureus |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.4.22.70 | recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Staphylococcus aureus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.4.22.70 | C184A | site-directed mutagenesis | Staphylococcus aureus |
| 3.4.22.70 | C184Hcy | site-directed mutagenesis, generation of a sortase mutant with Cys184 replaced by homocysteine (Hcy). Mutant Hcy-sortase is a poor catalyst with less than 1% of wild-type activity. The sensitivity of the active site nucleophiles towards an alkylation reagent correlates with the pKa values of the mutated residues | Staphylococcus aureus |
| 3.4.22.70 | C184Sec | site-directed mutagenesis, generation of a sortase mutant with Cys184 replaced by selenocysteine (Sec). Mutant Sec-sortase shows a moderate 2-3fold reduction in catalytic activity. The sensitivity of the active site nucleophiles towards an alkylation reagent correlates with the pKa values of the mutated residues. The pH-profile of mutant Sec-sortase is shifted to more acidic conditions when compared to the wild-type enzyme | Staphylococcus aureus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 3.4.22.70 | iodoacetamide | active site Cys184 of sortase A can be alkylated by iodoacetamide resulting in irreversible modified enzyme. The selenol and thiol of mutant Sec-sortase and mutant Hcy-sortase are sensitive to alkylation as well | Staphylococcus aureus |  |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.22.70 | additional information | Staphylococcus aureus | the bacterial transpeptidase sortase A catalyzes the chemoselective ligation of peptides and proteins. During catalysis sortase A cleaves the conserved Leu-Pro-X-Thr-Gly sorting motif at the Thr residue under concomitant thioester formation at active site Cys184 | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.4.22.70 | Staphylococcus aureus | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.4.22.70 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromaatography and dialysis | Staphylococcus aureus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.4.22.70 | Abz-LPKTGK(Dnp)KK + GGGWW | substrate peptide 1 (Abz-LPKTGK(Dnp)KK) and acceptor peptide 2 (GGGWW) | Staphylococcus aureus | Abz-LPKTGGGWW + GK(Dnp)KK | - |
? | |
| 3.4.22.70 | Dns-LPKTGGRR + GGGWW | dansyl-labeled Dns-LPKTGGRR substrate peptide and acceptor peptide 2 (GGGWW) | Staphylococcus aureus | Dns-LPKTGGGWW + GGRR | - |
? | |
| 3.4.22.70 | additional information | the bacterial transpeptidase sortase A catalyzes the chemoselective ligation of peptides and proteins. During catalysis sortase A cleaves the conserved Leu-Pro-X-Thr-Gly sorting motif at the Thr residue under concomitant thioester formation at active site Cys184 | Staphylococcus aureus | ? | - |
? | |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.70 | 25 | - |
assay at | Staphylococcus aureus |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.70 | 7.5 | - |
mutant Sec-sortase enzyme | Staphylococcus aureus |
| 3.4.22.70 | 8.5 | 9 | wild-type enzyme | Staphylococcus aureus |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.4.22.70 | 5.5 | 9.5 | mutant Sec-sortase enzyme, activity range, profile overview | Staphylococcus aureus |
| 3.4.22.70 | 6.5 | 11 | wild-type enzyme, activity range, profile overview | Staphylococcus aureus |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 3.4.22.70 | additional information | during catalysis sortase A cleaves the conserved Leu-Pro-X-Thr-Gly sorting motif at the Thr residue under concomitant thioester formation at active site Cys184, mechanism, overview | Staphylococcus aureus |
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