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

*この研究の対応する著者

研究成果: Article査読

15 被引用数 (Scopus)

抄録

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.

本文言語English
ページ(範囲)1272-1284
ページ数13
ジャーナルBiochimica et Biophysica Acta - Proteins and Proteomics
1804
6
DOI
出版ステータスPublished - 2010 6月
外部発表はい

ASJC Scopus subject areas

  • 分析化学
  • 生物理学
  • 生化学
  • 分子生物学

フィンガープリント

「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」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル