Calpain and calpastatin in rabbit corneal epithelium

Thomas (Tom) Shearer, Mitsuyoshi Azuma, Larry David, Yoko Yamagata, Takashi Murachi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The purpose of this study was to provide a direct assay for calpain and its endogenous inhibitor calpastatin in normal rabbit epithelium. Corneal epithelial extracts were fractionated by DEAE (1) chromatography on HPLC. Fractions were analyzed for calpain by ELISA, immunoblotting, and caseinolytic enzyme activity with FITC-labeled casein. Results demonstated Immunoreactive peaks for calpains I and II. Calpain II from the soluble fraction of corneal epithelium eluted at a similar NaCl concentration (260 raM) as calpain II from other tissues, was inhibited by both E64 and the removal of Ca, contained an 80 kDa subunit in immunoblots, and was present at specific activity of 220 units/g protein (in a crude homogenate). Calpain antigen was also present in the EDTA/EGTA washed insoluble fraction of corneal epithelium. Calpastatin in corneal epithelium eluted at 130 -160 mM NaCl on DEAE, coeluted with calpain I, and was present at 330 units/g protein (crude homogenate). The results demonstrated a calpain/calpastatin system in corneal epithelium, where it is speculated to play a role in epithelial cell turnover and wound healing.

Original languageEnglish (US)
Pages (from-to)39-44
Number of pages6
JournalCurrent Eye Research
Volume9
Issue number1
DOIs
StatePublished - 1990

Fingerprint

Corneal Epithelium
Calpain
Rabbits
calpastatin
Fluorescein-5-isothiocyanate
Egtazic Acid
Caseins
Immunoblotting
Edetic Acid
Wound Healing
Chromatography
Proteins
Epithelium
Epithelial Cells
Enzyme-Linked Immunosorbent Assay
High Pressure Liquid Chromatography
Antigens

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Calpain and calpastatin in rabbit corneal epithelium. / Shearer, Thomas (Tom); Azuma, Mitsuyoshi; David, Larry; Yamagata, Yoko; Murachi, Takashi.

In: Current Eye Research, Vol. 9, No. 1, 1990, p. 39-44.

Research output: Contribution to journalArticle

Shearer, Thomas (Tom) ; Azuma, Mitsuyoshi ; David, Larry ; Yamagata, Yoko ; Murachi, Takashi. / Calpain and calpastatin in rabbit corneal epithelium. In: Current Eye Research. 1990 ; Vol. 9, No. 1. pp. 39-44.
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