Is ubiquitination reversible?
Ubiquitination is a reversible process due to the presence of deubiquitinating enzymes that can cleave ubiquitin from modified proteins. Posttranslational modification of cell proteins, including ubiquitination, is involved in the regulation of both membrane trafficking and protein degradation.
How do you dilute MG132?
3 Prepare aliquots of MG-132 and store at -20 °C. 4. Once MG-132 has been dissolved, dilute 1 in 100 using sterile culture medium to obtain a solution at 200 µg/ml (420 µM).
What is the purpose of SUMOylation?
SUMOylation, as a post-translational modification, plays essential roles in various biological functions including cell growth, migration, cellular responses to stress and tumorigenesis. The imbalance of SUMOylation and deSUMOylation has been associated with the occurrence and progression of various diseases.
How do I get rid of SUMO tag?
SUMO fusion proteins can be cleaved to remove the SUMO moiety using SUMO-specific proteases such as Ulp1. Here, we describe the use of vectors for the expression of recombinant proteins in E. coli as fusions to the Drosophila SUMO protein.
What is the difference between CUL1 and CUL4?
CUL4 is a member of the cullin family of proteins, which share substantial homology to CUL1 originally identified in Caernorhabditis elegans [1].
What is the role of NEDD8 in CUL4 ligase activity?
CUL4 ligase activity is also regulated by the covalent attachment of the ubiquitin-like protein NEDD8 to CUL4, or neddylation, and the COP9 signalosome complex (CSN) that removes this important modification.
What is the function of neddylation in Cullin?
Cullin family proteins organize ubiquitin ligase (E3) complexes to target numerous cellular proteins for proteasomal degradation. Neddylation, the process that conjugates the ubiquitin-like polypeptide Nedd8 to the conserved lysines of cullins, is essential for in vivo cullin-organized E3 activities 1, 2.
What are the effects of targeted Neddylation in chronic lymphocytic leukemia (CLL)?
Targeting neddylation induces DNA damage and checkpoint activation and sensitizes chronic lymphocytic leukemia B cells to alkylating agents. Cell Death Dis.(2015) 6:e1807. 10.1038/cddis.2015.161 [PMC free article][PubMed] [CrossRef] [Google Scholar]