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J. Biochem, 2003, Vol. 134, No. 4 559-565
© 2003 Japanese Biochemical Society

CELL

Identification of the 58-kDa Phosphoprotein Associated with Motility Initiation of Hamster Spermatozoa

Masakatsu Fujinoki*,1, Takeshi Kawamura2, Toshifusa Toda3, Hideki Ohtake1, Nobuyoshi Shimizu2, Sadao Yamaoka1 and Makoto Okuno4

1 Department of Physiology, Dokkyo University School of Medicine, Mibu, Tochigi 321-0293; 2 Department of Molecular Biology, Keio University School of Medicine, Shinanomachi, Shinjuku-ku, Tokyo 160-8582; 3 TMIG Proteomics Collaboration Center, Tokyo Metropolitan Institute of Gerontology, Sakaecho, Itabashi-ku, Tokyo 173-0015; and 4 Department of Biology, Graduate School of Arts and Science, University of Tokyo, Komaba, Meguro-ku, Tokyo 153-8902

In our previous paper [M. Fujinoki et al. (2001) Biomed. Res. 22, 45–58], we reported that the 58-kDa protein obtained from hamster sperm flagella was phosphorylated at serine residues in association with the start of motility. In the present experiments, we identified and localized the 58-kDa protein. The 58-kDa protein was assumed to exist in the acrosomal region domain of the sperm head and the whole sperm flagellum. In particular, a large amount of 58-kDa protein was localized in the equatorial segment of the acrosomal region domain of the sperm head and the middle piece of the sperm flagellum. In the next step, the 58-kDa protein was identified by peptide mass finger printing and LC-MS/MS analysis. The results suggested that the 58-kDa protein was ATP synthase H+ transporting F1 ß, which is one of the mitochondrial components. Therefore, it is likely that the 58-kDa protein is associated with ATP production in the mitochondrial sheath in the middle piece of the sperm flagellum, and H+ transport in the sperm head and the sperm flagellum except for the middle piece, since ATP synthase also acts as an H+ pump.

* To whom correspondence should be addressed. Tel: +81-282-87-2125, Fax: +81-282-86-7835, E-mail: fujinoki{at}dokkyomed.ac.jp


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