Chemical proteomics profiling of proteasome activity.
Proteolysis is a key mechanism for protein homeostasis in living cells. This process is effected by different classes of proteases. The proteasome is one of the most abundant and versatile proteases, bearing three different proteolytic active sites. The proteasome plays an important role in essential biological pathways such as antigen presentation, signal transduction, and cell-cycle control feedback loops. The aim of this work is to design novel chemical strategies for capturing, detection, identification, and quantification--in one word, profiling--the active protease fractions of interest, in cells of different phenotypes. Here, a set of chemistry-based functional proteomics techniques is demonstrated by profiling the multi-catalytic protease activities of the proteasome. Importantly, functional profiling is complementary to expression level profiling and is an indispensable parameter for better understanding of mechanisms underlying biological processes.
Verdoes
Martijn
M
Leiden Institute of Chemistry, Leiden University, the Netherlands.
Berkers
Celia R
CR
Florea
Bogdan I
BI
van Swieten
Paul F
PF
Overkleeft
Herman S
HS
Ovaa
Huib
H
D015153:Blotting, Western; D002384:Catalysis; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D008956:Models, Chemical; D015345:Oligonucleotide Probes / Q000737:chemistry; D010447:Peptide Hydrolases / Q000737:chemistry; D010641:Phenotype; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction
D015345:Oligonucleotide Probes; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex