EC Number   |
Application |
Reference |
|---|
   3.4.22.68 | biotechnology |
substrate-trapping Ulp1(3)(C580S) interacts robustly with human SUMO1, SUMO2 and SUMO2 chains, making it a potentially useful tool for the analysis and purification of SUMO-modified proteins |
717436 |
| 3.4.22.68 | medicine |
there may be a connection between a defect in SUMO-1 conjugation to the PML protein and acute promyelocytic leukemia (ALP). Specific Ulp inhibitors can therefore have therapeutic value for ALP |
647396 |
| 3.4.22.68 | molecular biology |
modification of recombinant proteins by SUMOylation often dramatically increases solubility and stability during expression of the fusion proteins in bacteria relative to unfused proteins. After expressing a protein as a fusion to SUMO, it is often desirable to cleave the SUMO off of the fusion protein using a SUMO-specific protease such as Ulp1. To facilitate such processing, a dual expression vector is constructed encoding two fusion proteins: one consisting of SUMO fused to Ulp1 and a second consisting of SUMO fused to a His-tagged protein of interest. The SUMO-Ulp1 cleaves both itself and the other SUMO fusion protein in the bacterial cells prior to lysis, and the proteins retain solubility after cleavage, method evaluation, overview |
718357 |
| 3.4.22.68 | molecular biology |
usage of the targeting and small ubiquitin-like modifier, SUMO, binding properties of Ulp1(3)(C580S) to purify Smt3-modified proteins from cell extracts |
717436 |
| 3.4.22.68 | synthesis |
comparison of Ulp1 protease in active upon expression as inclusion bodies and soluble protein. Fusion of the N-terminal selfassembling peptide GFIL8 to Ulp1 increases production of active inclusion bodies in Escherichia coli. Attachment of the N-terminal cellulose-binding module facilitates the immobilization on regenerated amorphous cellulose with a binding capacity up to about 235 mg protein per gram of cellulose. The immobilized soluble Ulp1 maintains about 42% initial cleavage activity with repetitive use, whereas the aggregated Ulp1 loses its cleavage capacity after cleaving the protein substrate once. Crosslinking of inclusion bodies using glutaraldehyde inactivates Ulp1 |
754218 |
| 3.4.22.68 | synthesis |
immobilization of Ulp1 as a tool for cleavage of the SUMO tag of recombinant proteins. Ulp1 immobilized on N-hydroxysuccinimide-activated Sepharose maintains 95% substrate-cleavage ability and significantly enhanced pH and thermal stability. The immobilized Ulp1 can tolerate 15% (v/v) DMSO and 20% (v/v) ethanol. It can be reused for more than 15 batch reactions with 90% activity retention |
753054 |
| 3.4.22.68 | synthesis |
optimization of expression of the catalytic domain. Optimization of cultivation conditions at shake flask results in Ulp1 expression of 195 mg/l in TB medium. Ni-NTA affinity purification of Ulp1 using 0.1% Triton X-100, 0.01mM DTT, 0.02mM EDTA and 1% glycerol leads to a purity of about 95% with a recovery yield of 80% and specific activity of 398600 U/mg. The protease cleaves the SUMO tag even at 1:10,000 enzyme to substrate ratio The in vivo cleavage of SUMO tag via coexpression strategy also results in more than 80% cleavage of SUMO fusion protein |
753452 |
