Characterization of proteins involved in the regulation of sperm motility and their protein network

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The research was carried out to elucidate the molecular network or cascade from the reception of signaling molecule to the activation of motile machinery in sperm cell. As a result a part of the proteins involved in this signaling pathway have been identified and the molecular interactions among the molecules have been revealed. In the research period twelve proteins that undergo changes in electric point or in the amount were detected in the sperm of the ascidian Ciona intestinalis on two-dimentional gel electrophoresis and peptide mass fingerprinting, Among them, potentially important axonemal proteins could be identified : outer arm dynein intermediate chain2(IC2), an intermediate chain of inner arm dynein termed IC116, radial spoke component LRR37, the regulatory subunit of cAMP-dependent protein kinase and a 94 kDa novel armadillo repeat protein, Another proteins are though to be involved in the reception of signals or intracellular signal transduction, such as SP17-like protein, TER ATPasae,TEFB,and 14-3-3 proteins. On the other hand, protein-protein interaction was examined by means of selective extraction of the axonemes, a series of columm chromatography and immunoprecipitation. As a result, a subcomplex constitution the radial spoke could be isolated. The subcomptex was composed of AKAP-like protein (RSP3), HSP40, two MORN repeat proteins, RSP4/6 and several novel proteins whose homologs are also present in human but the functions have been unrevealed. The molecular interaction in this subcomplex would shed light on the molecular function of radial spoke and ultimately on the mechanism of flagellar motility conserved from ascidian to human. The result also suggests that molecular chaperone is involved in the formation, maintenance or regulation of flagellar motility.