Cloned (Comment) | Organism |
---|---|
gene ysaN, recombinant expression of N-terminally His-tagged wild-type and of His-tagged truncation mutant enzymes in Escherichia coli strain BL21(DE3), expression of untagged wild-type enzyme | Yersinia enterocolitica |
Protein Variants | Comment | Organism |
---|---|---|
additional information | generation of several truncation mutants of YsaN for identification of critical residues of YsaN for stable YsaL-YsaN complex formation, overview. Crosslinking of purified His-tagged enzyme YsaN and His-tagged regulator YsaL, pH 8.0, 25°C, using 0.5 mM ethylene glycol bis(sulfosuccinimidylsuccinate) and 1.5% glutaraldehyde, respectively | Yersinia enterocolitica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
hypothetical protein YE3555 termed YsaL | identification and evaluation of a negative regulator of YsaN, a hypothetical protein YE3555, termed YsaL. Purified YsaL is dimeric in solution and strongly associates with YsaN to form a stable heterotrimeric YsaL-YsaN complex with a stoichiometry of 2:1. The N terminal 6-20 residues of YsaN are invariably required for stable YsaL-YsaN complex formation. YsaL inhibits the ATPase activity of YsaN with a maximum inhibition at the molar ratio 2:1 (YsaL: YsaN). Protein YE3555 has a close evolutionary relationship with the T3S ATPase regulators. Identification of critical residues of YsaN for stable YsaL-YsaN complex formation, overview | Yersinia enterocolitica | |
NaCl | about 78% reduced activity at 500 mM NaCl | Yersinia enterocolitica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Yersinia enterocolitica |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
49500 | - |
12 * 49500, recombinant His-tagged enzyme, SDS-PAGE. The enzyme exists in solution as higher order oligomer. The ATPase activity of oligomeric YsaN is several fold higher than the activity of the monomeric form | Yersinia enterocolitica |
603000 | - |
recombinant His-tagged enzyme dodecamer, gel filtration | Yersinia enterocolitica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | Yersinia enterocolitica | - |
ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Yersinia enterocolitica | O30438 | gene ysaN | - |
Purification (Comment) | Organism |
---|---|
soluble part of recombinant N-terminally His-tagged wild-type enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration, recombinant His-tagged truncation mutant enzymes from Escherichia coli strain BL21(DE3) are unfolded, dialysed and refolded and purified by nickel affinity chromatography and gel filtration, recombinant untagged wild-type enzyme | Yersinia enterocolitica |
Renatured (Comment) | Organism |
---|---|
recombinant His-tagged truncation mutant enzymes from Escherichia coli strain BL21(DE3) are unfolded under denaturing conditions in 6 M guanidium-HCl, 25 mM Tris pH 8.0, 1 mM EDTA, 300 mM NaCl, followed by dialysis and refolding in 25 mM Tris, 50 mM NaCl | Yersinia enterocolitica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | - |
Yersinia enterocolitica | ADP + phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dodecamer | 12 * 49500, recombinant His-tagged enzyme, SDS-PAGE. The enzyme exists in solution as higher order oligomer. The ATPase activity of oligomeric YsaN is several fold higher than the activity of the monomeric form | Yersinia enterocolitica |
Synonyms | Comment | Organism |
---|---|---|
T3S ATPase | - |
Yersinia enterocolitica |
Type Three Secretion ATPase | - |
Yersinia enterocolitica |
YsaN ATPase | - |
Yersinia enterocolitica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Yersinia enterocolitica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Yersinia enterocolitica |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme YsaN is negatively regulated by YsaL, overview | Yersinia enterocolitica |