A class of protease for which the active sites include two histidine residues that coordinate a zinc ion. During catalysis, the Zn2+ promotes attack of the peptide carbonyl carbon by the oxygen atom of a water
RIPA lysis buffer supplemented with protease inhibitors was used to extract the whole cell lysates. The protein concentration of each sample was determined by the Bicinchoninic Acid Assay. Western blotting was performed as described in our previous studies [16]. The antibodies used in this study are...
MF: Molecular function OS: Overall survival PPI: Protein–protein interact TME: Tumor microenvironment TPM: Transcript per million TRIM8: Tripartite motif containing 8 UHRF2: Ubiquitin like with PHD and ring finger domains 2 USP: Ubiquitin-specific protease ...
In agreement with their dual Ub/SUMO protease activity, the XopD-like family, including XopD and examples from other plant-associated bacteria, are most closely related to the eukaryotic ULP families. As dedicated DUBs, the SseL-like examples (including ElaD and ShiCE) also segregate near the...
Cell pellets were lysed in lysis buffer (50 mm Tris, pH 7.5, 150 mm KCl, 0.5% CHAPS, 1 mm TCEP, 2 mm MgCl2, 1 mm phenylmethylsulfonyl fluoride, and protease inhibitors (Sigma)) for 1 h, and the lysate was cleared by centrifugation at 38,400 × g. The clarified lysate was ...
(DQ)-OVA was marginally reduced for cDC2, however, no striking differences were observed that sufficiently explained the reduction in antigen presentation (Fig.7d). Cathepsin protease activity was unaffected by the absence of UBL3 in cDC1 or cDC2 as profiled using an active site fluorescent ...
(Fig.1d). An additional Ser-Gly motif N-terminal of the first Pup residue is left over after TEV protease cleavage and is also resolved in the Mpa channel. The EM density of the engaged substrate becomes weaker following the Pup sequence out of the AAA chamber toward the double-OB ring ...
To test the interaction of Cdc48 with Cuz1, lysates were prepared by resuspending cell pellets in co-IP buffer A (25 mm Tris-HCl, pH 8.0, 200 mm NaCl, 2 mm MgCl2, 5% glycerol, 1% Triton X-100 and protease inhibitors) and, when indicated, 2 mm ATP. The resuspended cells were ...
As before, DNAse I and one protease inhibitor tablet were added to the solution, which was then rocked overnight. The resulting solution was clarified by ultracentrifugation and filtered. The supernatant was then loaded onto a 5 ml HisTrap (GE Healthcare) column pre-equilibrated with extraction ...
Cells were sonicated in the presence of phenylmethane sulfonyl fluoride and cOmplete EDTA-free (Roche) protease inhibitors and subsequently clarified by centrifugation. The protein was extracted from the solute by binding to Glutathione 4B Sepharose (GE) in batch. Resin was washed in sonication ...