Global profiling of protease cleavage sites by chemoselective labeling of protein N-termini.
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Xu G, Shin SB, Jaffrey SR
Global profiling of protease cleavage sites by chemoselective labeling of protein N-termini.
Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19310-5. doi: 10.1073/pnas.0908958106. Epub 2009 Nov 5.
- PubMed ID
- 19892738 [ View in PubMed]
- Abstract
Proteolysis has major roles in diverse biologic processes and regulates the activity, localization, and intracellular levels of proteins. Linking signaling pathways and physiologic processes to specific proteolytic processing events is a major challenge in signal transduction research. Here, we describe N-CLAP (N-terminalomics by chemical labeling of the alpha-amine of proteins), a general approach for profiling protein N-termini and identifying protein cleavage sites during cellular signaling. In N-CLAP, simple and readily available reagents are used to selectively affinity label the alpha-amine that characterizes the protein N terminus over the more highly abundant epsilon-amine on lysine residues. Protein cleavage sites are deduced by identifying the corresponding N-CLAP peptides, which are derived from the N-termini of proteins, including the N-termini of the newly formed polypeptide products of proteolytic cleavage. Through selective affinity purification and tandem mass spectrometry analysis of 278 N-CLAP peptides, we characterized proteolytic cleavage events associated with methionine aminopeptidases and signal peptide peptidases, as well as proteins that are proteolytically cleaved after cisplatin-induced apoptosis. Many of the protein cleavage sites that are elicited during apoptotic signaling are consistent with caspase-dependent cleavage. These data demonstrate the utility of N-CLAP for proteomic profiling of protein cleavage sites that are generated during cellular signaling.
DrugBank Data that Cites this Article
- Polypeptides
Name UniProt ID NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, mitochondrial O75489 Details NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mitochondrial O95178 Details Acyl carrier protein, mitochondrial O14561 Details Asparagine--tRNA ligase, cytoplasmic O43776 Details ATP-dependent Clp protease proteolytic subunit, mitochondrial Q16740 Details Ribosomal protein uL24-like Q9UNX3 Details ATP synthase subunit alpha, mitochondrial P25705 Details ATP synthase subunit beta, mitochondrial P06576 Details Cytochrome c oxidase subunit 6C P09669 Details Acireductone dioxygenase Q9BV57 Details ATP synthase subunit e, mitochondrial P56385 Details