Cloned (Comment) | Organism |
---|---|
the catalytic core domain of SENP1 (amino acids 415-643) and mutants are cloned into the vector pEHISTEV and expressed as a N-terminally His-tagged protein. The recombinant proteins are expressed in Escherichia coli Bl21(DE3) cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
SENP1 crystallization is performed at 20°C using a sitting drop vapour-diffusion method. Single diamond-shaped crystals are grown after 2 days from equal volumes of protein solution (20 mg/ml in 20 mM Tris/HCl, pH 8.0, and 50 mM NaCl) and reservoir solution containing 100 mM CoCl2, 0.1 M Mes, pH 6.5, and 1.8 M (NH4)2SO4. The structure of SENP1 is determined to 2.45 A. NaBH4 is used to trap a stable thiohemiacetal transition-state analogue between Cys602 of SENP1 and Gly92 of SUMO-2, determination of the structure of this complex to 3.2 A. Crystallization and structure determination of the SENP1-SUMO-2 complex | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C602A | completely inactive in processing of SUMO-2 and in deconjugating SUMO-2 | Homo sapiens |
D441A | mutant protein is indistinguishable from that of the wild-type protein in both processing and deconjugation | Homo sapiens |
D468A | mutated protein is only slightly impaired in processing and not at all in deconjugation | Homo sapiens |
D550A | completely inactive in processing of SUMO-2 and in deconjugating SUMO-2 | Homo sapiens |
F496A | altered protein retains significant levels of deconjugation activity | Homo sapiens |
H529A | altered protein retains significant levels of deconjugation activity | Homo sapiens |
H533A | completely inactive in processing of SUMO-2 and in deconjugating SUMO-2 | Homo sapiens |
additional information | residues 415643 of SENP1 are expressed and purified from Escherichia coli and are shown to be catalytically active in SUMO processing and deconjugation | Homo sapiens |
Q596A | mutant of SENP1 is severely impaired in both deconjugation and processing | Homo sapiens |
R511A | mutant has reduced activity, but is unaffected in deconjugation | Homo sapiens |
V532A | mutation has no effect on either deconjugation or processing | Homo sapiens |
W512A | mutant has a reduced deconjugation activity in vitro, its deconjugation activity in vivo is only impaired to a small extent | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q9P0U3 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(SUMO-1)-Ran GTPase-activating protein 1 conjugate + H2O | rates at which SUMO-1, SUMO-2 and SUMO-3 are deconjugated from RanGAP1 are indistinguishable | Homo sapiens | SUMO-1 + Ran GTPase-activating protein 1 | - |
? | |
(SUMO-2)-Ran GTPase-activating protein 1 conjugate + H2O | rates at which SUMO-1, SUMO-2 and SUMO-3 are deconjugated from RanGAP1 are indistinguishable | Homo sapiens | SUMO-2 + Ran GTPase-activating protein 1 | - |
? | |
(SUMO-2)n-glutathione S-transferase-promyelocytic leukemia protein conjugate + H2O | glutathione S-transferase-promyelocytic leukemia protein bearing a polymeric chain of SUMO-2 | Homo sapiens | ? | - |
? | |
(SUMO-3)-Ran GTPase-activating protein 1 conjugate + H2O | rates at which SUMO-1, SUMO-2 and SUMO-3 are deconjugated from RanGAP1 are indistinguishable | Homo sapiens | SUMO-3 + Ran GTPase-activating protein 1 | - |
? | |
SUMO-1 precursor + H2O | pre-SUMO-1 is processed rapidly | Homo sapiens | SUMO-1 + His-Ser-Thr-Val | - |
? | |
SUMO-2 precursor + H2O | pre-SUMO-2 is processed slowly | Homo sapiens | SUMO-2 + Val-Tyr | - |
? | |
SUMO-3 precursor + H2O | pre-SUMO-3 is processed at an intermediate rate | Homo sapiens | SUMO-3 + Val-Pro-Glu-Ser-Ser-Leu-Ala-Gly-His-Ser-Phe | - |
? |
Synonyms | Comment | Organism |
---|---|---|
SENP1 | - |
Homo sapiens |
sentrin-specific protease 1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Homo sapiens |