Identification of putative in vivo substrates of calpain 3 by comparative proteomics of overexpressing transgenic and nontransgenic mice

Details

Serval ID
serval:BIB_7ECBF301CF45
Type
Article: article from journal or magazin.
Collection
Publications
Title
Identification of putative in vivo substrates of calpain 3 by comparative proteomics of overexpressing transgenic and nontransgenic mice
Journal
Proteomics
Author(s)
Cohen  N., Kudryashova  E., Kramerova  I., Anderson  L. V., Beckmann  J. S., Bushby  K., Spencer  M. J.
ISSN
1615-9853 (Print)
Publication state
Published
Issued date
11/2006
Volume
6
Number
22
Pages
6075-84
Notes
Comparative Study
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't --- Old month value: Nov
Abstract
Calpain 3 (CAPN3) is a calcium-dependent protease, mutations in which cause limb girdle muscular dystrophy type 2A. To explore the physiological function of CAPN3, we compared the proteomes of transgenic mice that overexpress CAPN3 (CAPN3 Tg) and their nontransgenic (non-Tg) counterparts. We first examined known muscular dystrophy-related proteins to determine if overexpression of CAPN3 results in a change in their distribution or concentration. This analysis did not identify any known muscular dystrophy proteins as substrates of CAPN3. Next, we used a proteomic approach to compare and identify differentially represented proteins in 2-DE of CAPN3 Tg and non-Tg mice. LC-MS/MS analysis led to the identification of ten possible substrates for CAPN3, classified into two major functional categories: metabolic and myofibrillar. Myosin light chain 1 (MLC1) was focused upon because our previous studies suggested a role for CAPN3 in sarcomere remodeling. In this study, CAPN3 was shown to proteolyze MLC1 in vitro. These studies are the first to identify possible substrates for CAPN3 in an in vivo system and support a role for CAPN3 in sarcomere remodeling by cleavage of myofibrillar proteins such as MLC1. In addition, these data also suggest a role for CAPN3 in mitochondrial protein turnover.
Keywords
Animals Calpain/genetics/*metabolism Electrophoresis, Gel, Two-Dimensional Mass Spectrometry/methods Membrane Proteins/metabolism Mice Mice, Transgenic Models, Biological Muscle Proteins/genetics/*metabolism Muscle, Skeletal/*metabolism Muscular Dystrophies/*enzymology/metabolism Myosin Light Chains/metabolism Proteomics/*methods Substrate Specificity
Pubmed
Web of science
Create date
25/01/2008 17:18
Last modification date
03/03/2018 18:42
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