Calpain-1 resident in lipid raft/caveolin-1 membrane microdomains plays a protective role in endothelial cells

Biochimie. 2017 Feb;133:20-27. doi: 10.1016/j.biochi.2016.12.002. Epub 2016 Dec 9.

Abstract

We are here reporting that calpain-1 is a constitutive component of a distinct lipid raft/caveolin-1 microdomain isolated from bEnd5 cells in association with endothelial nitric oxide synthase (eNOS) and heat shock protein 90 (HSP90). Perturbations in intracellular calcium concentration by Ca2+-ionophore A23187 or prolonged cell exposure to high glucose induce a significant decrease in the level of eNOS accompanied by a recruitment of additional HSP90 molecules at this site. In these conditions the cells are more resistant to cell death by Ca2+ overload. The decrease of eNOS has been due not only to its Ca2+-mediated release from the caveolin-1 aggregates but also to its digestion by calpain-1. The specific involvement of calpain-1 in digestion of eNOS is supported by the preventive effect of a synthetic calpain inhibitor (CI-2) and by the absence of calpain-2 and calpastatin in the caveolin-1 microdomain. These results suggest that the protein adjustments observed in lipid raft/caveolin-1 microdomains could be visualized as a process required to protect the cells against NO overproduction and aberrant calpain activation. Alterations in eNOS, calpain-1 and HSP90 levels have been observed in aorta of Zucker Diabetic Rats (ZDR). The loss of HSP90 occurring in these animals indicates an aberrant activation of calpain and thereby the transition from a physiological to a pathological cell condition.

Keywords: Ca(2+)-dependent proteolysis; Calpain; Diabetes; HSP90; eNOS; nNOS.

MeSH terms

  • Animals
  • Aorta / metabolism
  • Aorta / pathology
  • Calcium / metabolism
  • Calpain / chemistry
  • Calpain / metabolism*
  • Caveolin 1 / chemistry
  • Caveolin 1 / metabolism*
  • Diabetes Mellitus / genetics
  • Diabetes Mellitus / metabolism*
  • Diabetes Mellitus / pathology
  • Endothelial Cells / metabolism*
  • Endothelial Cells / pathology
  • HSP90 Heat-Shock Proteins / chemistry
  • HSP90 Heat-Shock Proteins / metabolism*
  • Humans
  • Membrane Microdomains / metabolism
  • Nitric Oxide / biosynthesis
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type I / chemistry
  • Nitric Oxide Synthase Type I / metabolism
  • Nitric Oxide Synthase Type III / chemistry
  • Nitric Oxide Synthase Type III / metabolism
  • Oligopeptides / pharmacology
  • Proteolysis
  • Rats

Substances

  • Caveolin 1
  • HSP90 Heat-Shock Proteins
  • Oligopeptides
  • Nitric Oxide
  • N-(3-N-(benzyloxycarbonyl)amino-1-carboxypropyl)leucyl-O-methyltyrosine N-methylamide
  • Nitric Oxide Synthase Type I
  • Nitric Oxide Synthase Type III
  • Calpain
  • Calcium