Proteome-wide Identification of HtrA2/Omi Substrates
- 1 February 2007
- journal article
- Published by American Chemical Society (ACS) in Journal of Proteome Research
- Vol. 6 (3), 1006-1015
- https://doi.org/10.1021/pr060510d
Abstract
To identify apoptotic targets of HtrA2/Omi, we purified recombinant HtrA2/Omi and its catalytically inactive S306A mutant. Lysates of human Jurkat T lymphocytes incubated with either wild-type recombinant HtrA2/Omi or the S306A mutant were screened using the gel-free COFRADIC approach that isolates peptides covering the N-terminal parts of proteins. Analysis of the 1162 proteins identified by mass spectrometry yielded 15 HtrA2/Omi substrates of potential physiological relevance together holding a total of 50 cleavage sites. Several processing events were validated by incubating purified recombinant HtrA2/Omi with in vitro translated substrates or with Jurkat cell lysates. In addition, the generated set of cleavage sites was used to assess the protein substrate specificity of HtrA2/Omi. Our results suggest that HtrA2/Omi has a rather narrow cleavage site preference and that cytoskeletal proteins are prime targets of this protease.Keywords
This publication has 42 references indexed in Scilit:
- Neuroprotective Role of the Reaper-Related Serine Protease HtrA2/Omi Revealed by Targeted Deletion in MiceMolecular and Cellular Biology, 2004
- Binding Specificity and Regulation of the Serine Protease and PDZ Domains of HtrA2/OmiJournal of Biological Chemistry, 2003
- Loss of Omi mitochondrial protease activity causes the neuromuscular disorder of mnd2 mutant miceNature, 2003
- The HtrA Family of ProteasesMolecular Cell, 2002
- Crystal structure of DegP (HtrA) reveals a new protease-chaperone machineNature, 2002
- Identification of Omi/HtrA2 as a Mitochondrial Apoptotic Serine Protease That Disrupts Inhibitor of Apoptosis Protein-Caspase InteractionJournal of Biological Chemistry, 2002
- The Serine Protease Omi/HtrA2 Regulates Apoptosis by Binding XIAP through a Reaper-like MotifJournal of Biological Chemistry, 2002
- The Escherichia coli heat shock protease HtrA participates in defense against oxidative stressMolecular Genetics and Genomics, 1999
- A Temperature-Dependent Switch from Chaperone to Protease in a Widely Conserved Heat Shock ProteinCell, 1999
- The HtrA (DegP) protein, essential for Escherichia coli survival at high temperatures, is an endopeptidaseJournal of Bacteriology, 1990