Cloned (Comment) | Organism |
---|---|
expressed as a fusion with an N-terminal His6 tag in Escherichia coli strain BL21 (DE3) | Saccharolobus solfataricus |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method. The crystal structure of SSO1404 is solved at 1.6 A resolution revealing the first ribonuclease with a ferredoxin-like fold | Saccharolobus solfataricus |
Protein Variants | Comment | Organism |
---|---|---|
D10A | inactive mutant enzyme | Saccharolobus solfataricus |
D13A | mutant enzyme shows wild-type level activity | Saccharolobus solfataricus |
D14A | mutant enzyme shows wild-type level activity | Saccharolobus solfataricus |
D65A | 2fold drop in activity compared to wild-type | Saccharolobus solfataricus |
F37A | inactive mutant enzyme | Saccharolobus solfataricus |
N18A | mutant enzyme shows wild-type level activity | Saccharolobus solfataricus |
Q33A | slightly reduced activity | Saccharolobus solfataricus |
R17A | very low activity | Saccharolobus solfataricus |
R19A | very low activity | Saccharolobus solfataricus |
R31A | inactive mutant enzyme | Saccharolobus solfataricus |
R67A | mutant enzyme shows wild-type level activity | Saccharolobus solfataricus |
S35A | slightly reduced activity | Saccharolobus solfataricus |
T12A | 2fold drop in activity compared to wild-type | Saccharolobus solfataricus |
Y34A | slightly reduced activity | Saccharolobus solfataricus |
Y9A | very low activity | Saccharolobus solfataricus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
K+ | 100 mM, required | Saccharolobus solfataricus | |
Mg2+ | 5 mM, required | Saccharolobus solfataricus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
11936 | - |
2 * 11936, calculated from sequence | Saccharolobus solfataricus |
23700 | - |
gel filtration | Saccharolobus solfataricus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharolobus solfataricus | Q97YC2 | - |
- |
Saccharolobus solfataricus P2 | Q97YC2 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharolobus solfataricus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
single-stranded RNA + H2O | preferentially cleaves single-stranded RNAs within U-rich regions. Most cleavage sites contained one or two U. It cleaves the phosphodiester linkage on the 3'-side and generates 5'-phosphate- and 3'-hydroxyl-terminated oligonucleotides. The enzyme cleaves these substrates only in close proximity to the 5'- or 3'-ends suggesting that it requires the presence of a free RNA end. Oligoribonucleotides as short as 10 nt can serve as SSO1404 substrates. Tyr-9, Asp-10, Arg-17, Arg-19, Arg-31, and Phe-37 are important for enzymatic activity. Asp-10 might be the principal catalytic residue. No cleavage of dsRNA substrates prepared by annealing ssRNA substrates. No nuclease activity against either of the DNA substrates. The 39 nucleotide ssRNA substrate AAAUACG-/-U-/-U-/-UUCUCCAUUGUCAUAUUGCGCAUAAGUUGA shows the highest activity among the ssRNA substrates tested | Saccharolobus solfataricus | ? | - |
? | |
single-stranded RNA + H2O | preferentially cleaves single-stranded RNAs within U-rich regions. Most cleavage sites contained one or two U. It cleaves the phosphodiester linkage on the 3'-side and generates 5'-phosphate- and 3'-hydroxyl-terminated oligonucleotides. The enzyme cleaves these substrates only in close proximity to the 5'- or 3'-ends suggesting that it requires the presence of a free RNA end. Oligoribonucleotides as short as 10 nt can serve as SSO1404 substrates. Tyr-9, Asp-10, Arg-17, Arg-19, Arg-31, and Phe-37 are important for enzymatic activity. Asp-10 might be the principal catalytic residue. No cleavage of dsRNA substrates prepared by annealing ssRNA substrates. No nuclease activity against either of the DNA substrates. The 39 nucleotide ssRNA substrate AAAUACG-/-U-/-U-/-UUCUCCAUUGUCAUAUUGCGCAUAAGUUGA shows the highest activity among the ssRNA substrates tested | Saccharolobus solfataricus P2 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 11936, calculated from sequence | Saccharolobus solfataricus |
Synonyms | Comment | Organism |
---|---|---|
SSO1404 | locus name | Saccharolobus solfataricus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | 9 | - |
Saccharolobus solfataricus |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 10 | - |
Saccharolobus solfataricus |
General Information | Comment | Organism |
---|---|---|
physiological function | the role in the CRISPR-mediated anti-phage defense might involve degradation of phage or cellular mRNAs | Saccharolobus solfataricus |