| Spermatogenesis is a series of highly ordered process which involves many genes expression under precise temporal and spatial regulation. To better understand the molecular mechanism involved, we venture on a study to elucidate the expression, structure and function of genes encoding specific sperm proteins during various stages of spermatogenesis. Sperm surface proteins are important in sperm motility, sperm-egg recognition or binding to the zona pellucida and plasmalemma of the ovum. In a previous study, a novel membrane protein of human sperm, designated as YWK-II was isolated and characterized. Its open reading frame consists of 763 codons encoding a polypeptide characteristic of a single transmembrane protein established by hydropathicity analysis. Although the YWK-II gene has 70% homology to Alzheimer amyloid precursor protein (APP) within the transmembrane and cytoplasmic domains, the extracellular domain of YWK-II protein is distinctly different from that of APP and is shown to possess an anti-fertility activity. The preliminary results showed that the extracellular domain of YWK- II protein was able to interact with the 303-376 aminoacids segment of the rat Mullerian inhibiting substance (rMIS) in a yeast two-hybridsystem. We used molecular biological method to express, purify, crystallize two peptides of YWK-II from 436-674 and from 389-574 amino acids and solve the one of the extracellular fragments of YWK-II protein/APPH and obtained the complex crystal ofYWK-H and MIS.We also have captured the primary spermatocytes and round spermatids from rat testis sections by the application of the Laser Capture Microdissection (LCM) technique. Using the cDNAs from primary spermatocytes and round spermatids, the cDNA subtractive libraries of these cells were constructed by the application of the technology of suppression subtractive hybridization (SSH).RSB-66 was obtained from the SSH library of round spermatid-specific cDNAs against those of primary spermatocyte.
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