Altered calcium pump and secondary deficiency of gamma-sarcoglycan and microspan in sarcoplasmic reticulum membranes isolated from delta-sarcoglycan knockout mice

Cell Calcium. 2010 Jul;48(1):28-36. doi: 10.1016/j.ceca.2010.06.003. Epub 2010 Jul 16.

Abstract

Sarcoglycans (SGs) and sarcospan (SSPN) are transmembrane proteins of the dystrophin-glycoprotein complex. Mutations in the genes encoding SGs cause many inherited forms of muscular dystrophy. In this study, using purified membranes of wild-type (WT) and delta-SG knockout (KO) mice, we found the specific localization of the SG-SSPN isoforms in transverse tubules (TT) and sarcoplasmic reticulum (SR) membranes. Immunoblotting revealed that the absence of delta-SG isoforms in TT and SR results in a secondary deficiency of gamma-SG and microSPN. Our results showed augmented ATP hydrolytic activity, ATP-dependent calcium uptake and passive calcium efflux, probably through SERCA1 in KO compared to WT mice. Furthermore, we found a conformational change in SERCA1 isolated from KO muscle as demonstrated by calorimetric analysis. Following these alterations with mechanical properties, we found an increase in force in KO muscle with the same rate of fatigue but with a decreased fatigue recovery compared to WT. Together our observations suggest, for the first time, that the delta-SG isoforms may stabilize the expression of gamma-SG and microSPN in the TT and SR membranes and that this possible complex may play a role in the maintenance of a stable level of resting cytosolic calcium concentration in skeletal muscle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carrier Proteins / genetics*
  • Membrane Proteins / genetics*
  • Mice
  • Mice, Knockout
  • Muscle Fatigue / physiology
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / ultrastructure
  • Neoplasm Proteins / genetics*
  • Protein Isoforms / genetics
  • Sarcoglycans / deficiency*
  • Sarcoglycans / genetics
  • Sarcoplasmic Reticulum / genetics*
  • Sarcoplasmic Reticulum / metabolism
  • Sarcoplasmic Reticulum / ultrastructure
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / genetics*
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism

Substances

  • Atp2a1 protein, mouse
  • Carrier Proteins
  • Membrane Proteins
  • Neoplasm Proteins
  • Protein Isoforms
  • Sarcoglycans
  • Sspn protein, mouse
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases