Protein disulfide isomerase-P5, down-regulated in the final stage of boar epididymal sperm maturation, catalyzes disulfide formation to inhibit protein function in oxidative refolding of reduced denatured lysozyme

Kuniko Akama, Tomoe Horikoshi, Atsushi Sugiyama, Satoko Nakahata, Aoi Akitsu, Nobuyoshi Niwa, Atsushi Intoh, Yasutaka Kakui, Michiko Sugaya, Kazuo Takei, Noriaki Imaizumi, Takaya Sato, Rena Matsumoto, Hitoshi Iwahashi, Shin ichi Kashiwabara, Tadashi Baba, Megumi Nakamura, Tosifusa Toda

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

In mammalian spermiogenesis, sperm mature during epididymal transit to get fertility. The pig sharing many physiological similarities with humans is considered a promising animal model in medicine. We examined the expression profiles of proteins from boar epididymal caput, corpus, and cauda sperm by two-dimensional gel electrophoresis and peptide mass fingerprinting. Our results indicated that protein disulfide isomerase-P5 (PDI-P5) human homolog was down-regulated from the epididymal corpus to cauda sperm, in contrast to the constant expression of protein disulfide isomerase A3 (PDIA3) human homolog. To examine the functions of PDIA3 and PDI-P5, we cloned and sequenced cDNAs of pig PDIA3 and PDI-P5 protein precursors. Each recombinant pig mature PDIA3 and PDI-P5 expressed in Escherichia coli showed thiol-dependent disulfide reductase activities in insulin turbidity assay. Although PDIA3 showed chaperone activity to promote oxidative refolding of reduced denatured lysozyme, PDI-P5 exhibited anti-chaperone activity to inhibit oxidative refolding of lysozyme at an equimolar ratio. SDS-PAGE and Western blotting analysis suggested that disulfide cross-linked and non-productively folded lysozyme was responsible for the anti-chaperone activity of PDI-P5. These results provide a molecular basis and insights into the physiological roles of PDIA3 and PDI-P5 in sperm maturation and fertilization.

Original languageEnglish
Pages (from-to)1272-1284
Number of pages13
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1804
Issue number6
DOIs
Publication statusPublished - 2010 Jun
Externally publishedYes

Keywords

  • Anti-chaperone activity
  • Protein disulfide isomerase A3
  • Protein disulfide isomerase-P5
  • Proteomics
  • Sperm maturation
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biophysics
  • Biochemistry
  • Molecular Biology

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    Akama, K., Horikoshi, T., Sugiyama, A., Nakahata, S., Akitsu, A., Niwa, N., Intoh, A., Kakui, Y., Sugaya, M., Takei, K., Imaizumi, N., Sato, T., Matsumoto, R., Iwahashi, H., Kashiwabara, S. I., Baba, T., Nakamura, M., & Toda, T. (2010). Protein disulfide isomerase-P5, down-regulated in the final stage of boar epididymal sperm maturation, catalyzes disulfide formation to inhibit protein function in oxidative refolding of reduced denatured lysozyme. Biochimica et Biophysica Acta - Proteins and Proteomics, 1804(6), 1272-1284. https://doi.org/10.1016/j.bbapap.2010.02.004