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A proteomic study of calpain-3 and its involvement in limb girdle muscular dystrophy type 2A.

Bertipaglia, I; Richard, I; Påhlman, Anna-Karin LU ; Andersson, Liselotte LU ; James, Peter LU and Carafoli, E (2009) In Cell Calcium 46(5-6). p.356-363
Abstract (Swedish)
Abstract in Undetermined

Limb-girdle muscular dystrophy type 2A is an autosomal recessive disorder generated by inactivating mutations in the gene coding for the muscle specific protease calpain-3. It is mainly expressed in skeletal muscle as a monomeric multidomain protein characterized by three unique insertion sequences (NS, IS1, IS2). It is unstable, and undergoes very rapid autolysis in solution, therefore, its heterologous expression and purification have been difficult. So far, calpain-3 substrates have been only identified in vitro and with indirect approaches. We have therefore decided to perform a comprehensive study of the substrates of the protease by comparing the 2D electrophoretic profile of myotubes from... (More)
Abstract in Undetermined

Limb-girdle muscular dystrophy type 2A is an autosomal recessive disorder generated by inactivating mutations in the gene coding for the muscle specific protease calpain-3. It is mainly expressed in skeletal muscle as a monomeric multidomain protein characterized by three unique insertion sequences (NS, IS1, IS2). It is unstable, and undergoes very rapid autolysis in solution, therefore, its heterologous expression and purification have been difficult. So far, calpain-3 substrates have been only identified in vitro and with indirect approaches. We have therefore decided to perform a comprehensive study of the substrates of the protease by comparing the 2D electrophoretic profile of myotubes from obtained from calpain-3 knockout and wild type mice. Digestion of differentially expressed spots was followed by mass spectrometry analysis. We could identify 16 proteins which differed in knockout and wild type mice. Among them: desmin, nestin, spectrin and PDLIM1 were of particular interest. In vitro experiments have then revealed that only PDLIM1 is cleaved directly by the protease, and that a fragment of about 8 kDa is released from the C-terminal portion of the protein. (C) 2009 Elsevier Ltd. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Calpain, Muscular dystrophy, Proteomics, PDLIM 1
in
Cell Calcium
volume
46
issue
5-6
pages
356 - 363
publisher
Elsevier
external identifiers
  • wos:000273411200006
  • scopus:70649088773
ISSN
0143-4160
DOI
10.1016/j.ceca.2009.10.003
language
English
LU publication?
yes
id
f08088b5-be87-4902-a15b-d81e5b35937e (old id 1394514)
date added to LUP
2009-05-12 12:15:33
date last changed
2017-01-01 06:35:29
@article{f08088b5-be87-4902-a15b-d81e5b35937e,
  abstract     = {<b>Abstract in Undetermined</b><br/><br>
Limb-girdle muscular dystrophy type 2A is an autosomal recessive disorder generated by inactivating mutations in the gene coding for the muscle specific protease calpain-3. It is mainly expressed in skeletal muscle as a monomeric multidomain protein characterized by three unique insertion sequences (NS, IS1, IS2). It is unstable, and undergoes very rapid autolysis in solution, therefore, its heterologous expression and purification have been difficult. So far, calpain-3 substrates have been only identified in vitro and with indirect approaches. We have therefore decided to perform a comprehensive study of the substrates of the protease by comparing the 2D electrophoretic profile of myotubes from obtained from calpain-3 knockout and wild type mice. Digestion of differentially expressed spots was followed by mass spectrometry analysis. We could identify 16 proteins which differed in knockout and wild type mice. Among them: desmin, nestin, spectrin and PDLIM1 were of particular interest. In vitro experiments have then revealed that only PDLIM1 is cleaved directly by the protease, and that a fragment of about 8 kDa is released from the C-terminal portion of the protein. (C) 2009 Elsevier Ltd. All rights reserved.},
  author       = {Bertipaglia, I and Richard, I and Påhlman, Anna-Karin and Andersson, Liselotte and James, Peter and Carafoli, E},
  issn         = {0143-4160},
  keyword      = {Calpain,Muscular dystrophy,Proteomics,PDLIM 1},
  language     = {eng},
  number       = {5-6},
  pages        = {356--363},
  publisher    = {Elsevier},
  series       = {Cell Calcium},
  title        = {A proteomic study of calpain-3 and its involvement in limb girdle muscular dystrophy type 2A.},
  url          = {http://dx.doi.org/10.1016/j.ceca.2009.10.003},
  volume       = {46},
  year         = {2009},
}