PMID: 11331208May 2, 2001Paper

Identification of glycogen phosphorylase and creatine kinase as calpain substrates in skeletal muscle

The International Journal of Biochemistry & Cell Biology
J PurintrapibanN E Forsberg

Abstract

The goal of this study was to identify calpain substrates in muscle cells. Our hypothesis was that the yeast two-hybrid method could be used to identify novel calpain substrates. To accomplish this, native mu- and m-calpains, as well as a variety of calpain DNA fragments, were expressed in yeast cells and used to screen for binding proteins in a human skeletal muscle cDNA library. Calpain constructs that were used in the screening process included native mu- and m-calpains, a dominant negative (DN) m-calpain (i.e. active site modified), N-terminal truncated DN m-calpain (i.e. autolyzed DN-m-calpain) and, finally, an N- and C-terminal truncated m-calpain (i.e. autolyzed DN-m-calpain lacking a calcium-binding domain). Yeast cells were transformed using yeast two-hybrid expression vectors containing the different calpain constructs as "baits". Beta-galactosidase activity was assayed as an index of interaction between calpain and its potential target proteins. From this analysis, four clones (Ca2+-ATPase, novel nebulin-related protein (N-RAP), creatine kinase and glycogen phosphorylase) were recovered. Two of these, creatine kinase and glycogen phosphorylase, were selected for further study. In in-vitro assays, calpain was able to ...Continue Reading

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Citations

Oct 21, 2011·Journal of Agricultural and Food Chemistry·Phodchanee Phongpa-NganLouise Wicker
Aug 8, 2006·Critical Reviews in Food Science and Nutrition·Christine Delbarre-LadratVéronique Verrez-Bagnis
Sep 3, 2003·Bioscience, Biotechnology, and Biochemistry·Eiichi KimuraHiroyuki Sorimachi
Sep 11, 2002·Biochemical and Biophysical Research Communications·Rong GuoL Darryl Quarles
May 1, 2013·Expert Opinion on Therapeutic Patents·Nicolas Gaboriaud-Kolar, Alexios-Leandros Skaltsounis
Aug 18, 2016·Journal of Food Science·Wiesław PrzybylskiUrszula Jankiewicz
Sep 28, 2018·Medicinal Research Reviews·Yacoubou Abdoul Razak MahamanXiaochuan Wang
Mar 2, 2019·Animals : an Open Access Journal From MDPI·Martin ŠkrlepMarjeta Čandek-Potokar
Sep 19, 2002·Journal of Agricultural and Food Chemistry·René LametschEmøke Bendixen

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