Differential influence of proteolysis by calpain 2 and Lp82 on in vitro precipitation of mouse lens crystallins

Mitsuyoshi Azuma, Yoshiyuki Tamada, Sayaka Kanaami, Emi Nakajima, Yoshikuni Nakamura, Chiho Fukiage, Neil E. Forsberg, Melinda K. Duncan, Thomas R. Shearer

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

The purpose of the present study was to compare the susceptibility of crystallins proteolyzed by ubiquitous calpain 2 and by lens-specific calpain Lp82 to insolubilization. To test this, transgenic (TG) mice expressing a calpain 2, in which the active site cysteine 105 was mutated to alanine, were produced. Expression of mutated calpain 2 was driven in lens by coupling the mutated gene to the βB1-crystallin promoter. Light scattering was measured in solutions of lens proteins after activation of endogenous calpain 2 and/or Lp82. Mass spectrometric analysis was performed to determine the cleavage sites and the calpain responsible for insolubilization of crystallins. Lens proteins from TG mice incubated in vitro with calcium showed higher light scattering compared to proteins from wild type (WT) mice. αA-crystallin from TG mice was proteolyzed by Lp82. In contrast, αA-crystallin in lenses from WT mice were proteolyzed by both calpain 2 and Lp82. These results suggested that Lp82-induced proteolysis of crystallins caused increased susceptibility of truncated crystallins to in vitro precipitation. Since Lp82 is highest in young animals, Lp82-induced proteolysis and precipitation may be one of the factors responsible for the cataract formation in young rodents.

Original languageEnglish (US)
Pages (from-to)558-563
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume307
Issue number3
DOIs
StatePublished - Aug 1 2003

Keywords

  • Calpain 2
  • Crystallin
  • Insolubilization
  • Light scattering
  • Lp82
  • Mass spectrometry
  • Transgenic mouse

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Azuma, M., Tamada, Y., Kanaami, S., Nakajima, E., Nakamura, Y., Fukiage, C., Forsberg, N. E., Duncan, M. K., & Shearer, T. R. (2003). Differential influence of proteolysis by calpain 2 and Lp82 on in vitro precipitation of mouse lens crystallins. Biochemical and Biophysical Research Communications, 307(3), 558-563. https://doi.org/10.1016/S0006-291X(03)01194-X