Comprehensive survey of p94/calpain 3 substrates by comparative proteomics – Possible regulation of protein synthesis by p94
Open Access
- 4 May 2007
- journal article
- research article
- Published by Wiley in Biotechnology Journal
- Vol. 2 (5), 565-576
- https://doi.org/10.1002/biot.200700018
Abstract
Calpain represents a family of Ca2+-dependent cytosolic cysteine proteases found in almost all eukaryotes and some bacteria, and is involved in a variety of biological phenomena, including brain function. Several substrates of calpain are aggressively proteolyzed under pathological conditions, e.g., in neurodegenerating processes, fodrin is proteolyzed by calpain. Because very small amounts of substrate are proteolyzed by calpain under normal biological conditions, the molecular identities of calpain substrates are largely unknown. In this study, an extensive survey of the substrates of p94/calpain 3 in COS7 cells was executed using iTRAQTM labeling and 2-D LC-MALDI analysis. p94 was used because: (i) several p94 splicing variants are expressed in brain tissue even though p94 itself is a skeletal-muscle-specific calpain, and (ii) it exhibits Ca2+-independent activity in COS cells, which makes it useful for evaluating the effects of p94 protease activity on proteins without perturbing the cells. Our approach revealed several novel protein substrates for p94, including the substrates of conventional calpains, components of the protein synthesis system, and enzymes of the glycolytic pathway. The results demonstrate the usefulness and sensitivity of this approach for mining calpain substrates. A combination of this method with other analytical methods would contribute to elucidation of the biological relevance of the calpain family.Keywords
This publication has 48 references indexed in Scilit:
- Molecular and cellular basis of calpainopathy (limb girdle muscular dystrophy type 2A)Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2007
- Identification of putativein vivo substrates of calpain 3 by comparative proteomics of overexpressing transgenic and nontransgenic miceProteomics, 2006
- Regulation of the M-Cadherin-β-Catenin Complex by Calpain 3 during Terminal Stages of Myogenic DifferentiationMolecular and Cellular Biology, 2006
- Calpain 3: a key regulator of the sarcomere?The FEBS Journal, 2006
- Lysosomal biogenesis and function is critical for necrotic cell death in Caenorhabditis elegans The Journal of cell biology, 2006
- Multiplexed Protein Quantitation in Saccharomyces cerevisiae Using Amine-reactive Isobaric Tagging ReagentsMolecular & Cellular Proteomics, 2004
- Newly identified exons encoding novel variants of p94/calpain 3 are expressed ubiquitously and overlap the α‐glucosidase C geneFEBS Letters, 2003
- Characterization of a new p94-like calpain form in human lymphocytesBiochemical Journal, 2003
- Disruption of the Murine Calpain Small Subunit Gene, Capn4: Calpain Is Essential for Embryonic Development but Not for Cell Growth and DivisionMolecular and Cellular Biology, 2000
- Human endothelial actin-binding protein (ABP-280, nonmuscle filamin): a molecular leaf spring.The Journal of cell biology, 1990