Cloned (Comment) | Organism |
---|---|
gene cshA, recombinant expression of His-tagged enzyme fragment 1-211, comprising the N-terminal RecA-like domain 1, in Escherichia coli strain Rosetta2 (DE3) | Staphylococcus aureus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant N-terminal RecA-like domain 1 of DEAD-box RNA helicase CshA free and in complex with AMP or AMP-PNP, sitting drop vapor diffusion method, mixing of 0.001 ml of 5 mg/ml protein in 50 mM Tris-HCl, pH 7.5, 500 mM NaCl, with AMP or AMP-PNP in a molar ratio of 1:5 for the complexes, is mixed with 0.001 ml of reservoir solution containing 0.2 M ammonium acetate, 0.1 M sodium citrate tribasic dihydrate, pH 5.6, 30% v/v PEG 4000, at 16°C, X-ray diffraction structure determination and analysis at 1.5 and 1.8 A resolution, respectively, molecular replacement modelling | Staphylococcus aureus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | requird | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | Staphylococcus aureus | - |
ADP + phosphate | - |
? | |
ATP + H2O | Staphylococcus aureus ATCC 700699 | - |
ADP + phosphate | - |
? | |
ATP + H2O | Staphylococcus aureus Mu50 | - |
ADP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | Q99SH6 | - |
- |
Staphylococcus aureus ATCC 700699 | Q99SH6 | - |
- |
Staphylococcus aureus Mu50 | Q99SH6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme N-terminal RecA-like domain 1 from Escherichia coli strain Rosetta2 (DE3) by nickel affinity chromatography, gel filtration, and ultrafiltration | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + H2O | - |
Staphylococcus aureus | ADP + phosphate | - |
? | |
ATP + H2O | - |
Staphylococcus aureus ATCC 700699 | ADP + phosphate | - |
? | |
ATP + H2O | - |
Staphylococcus aureus Mu50 | ADP + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CshA | - |
Staphylococcus aureus |
DEAD-box RNA helicase | - |
Staphylococcus aureus |
General Information | Comment | Organism |
---|---|---|
evolution | CshA is a DEAD-box RNA helicase that belongs to the DExD/H-box family of proteins, which generally have an RNA-dependent ATPase activity | Staphylococcus aureus |
additional information | the N-terminal RecA-like domain 1 (amino acids 1-211) of DEAD-box RNA helicase CshA adopts a conserved alpha/beta RecA-like structure with seven parallel strands surrounded by nine alpha-helices. The Q motif and motif I are responsible for the binding of the adenine group and phosphate group of AMP, respectively. Motif I undergoes a conformational change upon AMP binding. Essential roles of Phe22 in the Q motif and Lys52 in motif I for binding ATP, indicating a conserved substrate-binding mechanism in SaCshA compared with other DEAD-box RNA helicases. Structure comparisons, overview | Staphylococcus aureus |
physiological function | enzyme CshA is a component of the RNA degradosome and plays important roles in RNA turnover | Staphylococcus aureus |