老鼠貯精囊蛋白之研究

Abstract

使用膠體過瀘法，陽離子交換樹脂，吾人系統地分離了老鼠貯精囊蛋白，且進一步以逆轉相高壓液體層析儀而純化了SVSVII，其分子量約為一萬，但卻含有四對雙硫鍵，顯示其乃一十分推擠的蛋白，其氨基酸組成及氮端順序已被決定 。 SVSIV 乃一雄性荷爾蒙（androgen）的標的蛋白，業已分離完成，以之打入兔子體內而誘導出相當專一性的抗體，利用此抗體在各組識中篩選而證明SVSIV蛋白只存在於貯精囊中，以此資訊吾人可以此抗體為探針來追蹤貯精囊在分化及成長過程中雄性荷爾蒙所扮演的角色。 貯精囊分泌液在精液中約占百分之八十左右，初步研究對sperm motility並無顯著的影響，但有待進一步的研究。

SVS VII was purified by a combination of gel filtration, ion exchange chromatography, and reverse phase HPLC to homogeneity as tested by NaDodSO4 polyacrylamide gel electrophoresis. The purified protein has a molecular weight of 9,000-10,000 dalton with eight cysteine residues. It is likely to be a very compact protein. Its N termimal sequence was determined as Leu-Ile-???-Asn-Ser-Val-Gly-Lys-Ser. The sequence of peptide segments obtained from digestion of the protein with CNBr was determined as Leu-Ile-Pro-Asn(Lys)-Glu-Val(Leu)-Ala(Glu)-Lys-Pro-Gly-Asp(Glu)-Ser-Gly-Lys-Thr-Val and Glu(Asp)-Ser-Pro-Glu-Glu-Lys-Glu-Lys-Glu-Leu-Asn. Antiserum against SVS IV was raised in rabbits and the specific antibody was purified. The purified antibody was used to survey the SVS IV distribution in different organs. We demonstrated that the SVS IV was only present in the seminal vesicle. No significant effect of seminal vesicle fluid on the sperm motility was found.