Lp82 is the dominant form of calpain in young mouse lens

Hong Ma, I. Hata, M. Shih, C. Fukiage, Y. Nakamura, M. Azuma, Thomas (Tom) Shearer

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The purpose of this study was to characterize Lp82 calpain in normal mouse. Lp82 is a lens-specific, calcium-activated isozyme from the calpain super family of cysteine proteases (EC 34.22.17). RT-PCR and molecular cloning were performed on total RNA from 12 day-old mice. Lp82 and m-calpain protein levels and proteolytic activities in lenses were measured by casein zymography, immunoblotting, and ELISA after partial purification by DEAE- HPLC. The 2334-bp cDNA encoding for mouse Lp82 contained a single large open reading frame encoding a protein of 709 amino acid residues with a calculated molecular weight of 82.2 kDa and a predicted pI of 5.8. The amino acid sequence of mouse lens Lp82 was 99 % homologous to rat lens Lp82. As in rat, mouse lens Lp82 showed a unique N-terminus and deletion of the IS1 and IS2 regions. In contrast to rat, Lp82 was the dominant calpain in young mouse lens. Lp82 was lens-specific, and the lens nucleus contained the highest specific activity of Lp82 and very little m-calpain. Endogenous Lp82 in lens soluble proteins was activated by addition of calcium and caused limited proteolysis of crystallins even in the presence of large amounts of recombinant domain I from the natural calpain inhibitor calpastatin. Loss of Lp82 protein accompanied aging of mouse lens. Lp82 may be responsible for a major portion of crystallin proteolysis occurring during normal lens development and maturation, or during cataract formation in young mice.

Original languageEnglish (US)
Pages (from-to)447-456
Number of pages10
JournalExperimental Eye Research
Volume68
Issue number4
DOIs
StatePublished - Apr 1999

Fingerprint

Calpain
Lenses
Crystallins
Proteolysis
Calcium
Proteins
Cysteine Proteases
Molecular Cloning
Caseins
Immunoblotting
Cataract
Open Reading Frames
Isoenzymes
Amino Acid Sequence
Complementary DNA
Molecular Weight
Enzyme-Linked Immunosorbent Assay
High Pressure Liquid Chromatography
RNA
Amino Acids

Keywords

  • Casein zymography
  • Immunoblot
  • Lens maturation
  • Lp82
  • M-calpain
  • Molecular cloning
  • Mouse lens
  • mRNA

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Ma, H., Hata, I., Shih, M., Fukiage, C., Nakamura, Y., Azuma, M., & Shearer, T. T. (1999). Lp82 is the dominant form of calpain in young mouse lens. Experimental Eye Research, 68(4), 447-456. https://doi.org/10.1006/exer.1998.0625

Lp82 is the dominant form of calpain in young mouse lens. / Ma, Hong; Hata, I.; Shih, M.; Fukiage, C.; Nakamura, Y.; Azuma, M.; Shearer, Thomas (Tom).

In: Experimental Eye Research, Vol. 68, No. 4, 04.1999, p. 447-456.

Research output: Contribution to journalArticle

Ma, H, Hata, I, Shih, M, Fukiage, C, Nakamura, Y, Azuma, M & Shearer, TT 1999, 'Lp82 is the dominant form of calpain in young mouse lens', Experimental Eye Research, vol. 68, no. 4, pp. 447-456. https://doi.org/10.1006/exer.1998.0625
Ma H, Hata I, Shih M, Fukiage C, Nakamura Y, Azuma M et al. Lp82 is the dominant form of calpain in young mouse lens. Experimental Eye Research. 1999 Apr;68(4):447-456. https://doi.org/10.1006/exer.1998.0625
Ma, Hong ; Hata, I. ; Shih, M. ; Fukiage, C. ; Nakamura, Y. ; Azuma, M. ; Shearer, Thomas (Tom). / Lp82 is the dominant form of calpain in young mouse lens. In: Experimental Eye Research. 1999 ; Vol. 68, No. 4. pp. 447-456.
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