Molecular cloning and characterization of functional domains of a human testis-specific isoform of calpastatin

Siming Li, Zhi Guo Liang, Gui Yu Wang, Bella Yavetz, Edward Kim, Erwin Goldberg

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Human serum containing sperm-agglutinating antibodies was used to screen a testis cDNA expression library to identify the cognate antigens that may be responsible for this biological effect. The longest positive phage clone (1.9 kb) was sequenced and found to be a testis-specific isoform of calpastatin (tCAST). The testis-specific segment of tCAST is encoded by a single exon within intron 14 of the calpastatin gene. A unique protein isoform is produced that differs in domain structure from the somatic calpastatins (sCAST). Human sCAST most commonly has an N-terminal domain L plus the four functional calpain inhibitory domains. Human tCAST consists of a 40-amino- acid N-terminal T domain plus a part of domain II and all of domains III and IV from the somatic isoform. Our data show that the T domain can target cytosolic localization and membrane association of tCAST, whereas domain I of sCAST exhibits a nuclear localization function. Calpastatin is the endogenous inhibitor of calpain. The calpain/calpastatin system is involved in membrane fusion events for several cell types, and calpain has been localized to the sperm acrosome. We detected tCAST in human sperm and testes extracts by Western blotting with specific antisera. These observations suggest that tCAST may modulate calpain in the calcium-mediated acrosome reaction that is required for fertilization.

Original languageEnglish (US)
Pages (from-to)172-178
Number of pages7
JournalBiology of Reproduction
Volume63
Issue number1
DOIs
StatePublished - Jan 1 2000
Externally publishedYes

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Molecular Cloning
Testis
Protein Isoforms
Calpain
Spermatozoa
calpastatin
Acrosome Reaction
Acrosome
Membrane Fusion
Gene Library
Fertilization
Bacteriophages
Introns
Immune Sera
Exons
Clone Cells
Western Blotting
Calcium
Antigens
Amino Acids

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Molecular cloning and characterization of functional domains of a human testis-specific isoform of calpastatin. / Li, Siming; Liang, Zhi Guo; Wang, Gui Yu; Yavetz, Bella; Kim, Edward; Goldberg, Erwin.

In: Biology of Reproduction, Vol. 63, No. 1, 01.01.2000, p. 172-178.

Research output: Contribution to journalArticle

Li, Siming ; Liang, Zhi Guo ; Wang, Gui Yu ; Yavetz, Bella ; Kim, Edward ; Goldberg, Erwin. / Molecular cloning and characterization of functional domains of a human testis-specific isoform of calpastatin. In: Biology of Reproduction. 2000 ; Vol. 63, No. 1. pp. 172-178.
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