Alterations of protein degradation and 2-D protein pattern in muscle cells of MDX and DMD origin

Biochem Biophys Res Commun. 1992 Dec 30;189(3):1484-90. doi: 10.1016/0006-291x(92)90242-d.

Abstract

Intracellular protein turnover of MDX, DMD and normal muscle was determined by [35S]methionine pulse-chase experiments and subsequent high resolution 2-D gel electrophoresis. In MDX myotubes intracellular degradation of short-lived and long-lived proteins was markedly increased by a factor of 1,4-2,1. In wildtype the rate of degradation of short-lived proteins was approximately 2.6%/h, whereas in MDX these proteins were degraded by 5.7%/h. Long-lived proteins were degraded in wildtype at a rate of 1.8%/h, and in MDX at a rate of 2.5%/h. Furthermore, we have described a 51.000 Da protein with an IEP of 5.1 (p51/5.1), whose net content is highly and specifically reduced in cultured MDX and DMD muscle cells as well as in isolated MDX muscle fibers. Treatment with calcium-channel blockers Dantrolene and Verapamil inhibited the degradation of p51/5.1 in MDX myotubes by more than 90% in contrast to controls.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / biosynthesis
  • Actins / isolation & purification
  • Actins / metabolism
  • Animals
  • Autoradiography
  • Cells, Cultured
  • Electrophoresis, Gel, Two-Dimensional
  • Kinetics
  • Methionine / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Mutant Strains
  • Muscle Proteins / biosynthesis
  • Muscle Proteins / isolation & purification
  • Muscle Proteins / metabolism*
  • Muscles / cytology
  • Muscles / metabolism*
  • Muscles / pathology
  • Muscular Dystrophy, Animal / metabolism*
  • Proton-Translocating ATPases / biosynthesis
  • Proton-Translocating ATPases / isolation & purification
  • Proton-Translocating ATPases / metabolism
  • Sulfur Radioisotopes
  • Tubulin / biosynthesis
  • Tubulin / isolation & purification
  • Tubulin / metabolism*
  • Vimentin / biosynthesis
  • Vimentin / isolation & purification
  • Vimentin / metabolism

Substances

  • Actins
  • Muscle Proteins
  • Sulfur Radioisotopes
  • Tubulin
  • Vimentin
  • Methionine
  • Proton-Translocating ATPases