Wang L F, Wei S G, Miao S Y, Liu Q Y, Koide S S
National Laboratory of Medical Molecular Biology, Beijing, China.
Biochem Mol Biol Int. 1994 May;33(2):245-51.
Antibodies present in a serum obtained from an infertile woman interacted with a 17.5 kD glycoprotein (BS-17 component) extracted from human sperm by Western blot. Polyclonal antibodies were raised against the BS-17 component and used to identify positive staining clones from a human testis lambda gt11 expression library. The cDNA encoding the BS-17 component was isolated and its nucleotide sequence determined. The BS-17 cDNA contained 758 nucleotides with an open reading frame of 558 nucleotides encoding a polypeptide consisting of 186 amino acids. The BS-17 cDNA showed 99.7% homology in 758 nucleotides overlap with the 3' terminus of the gene coding calpastatin and 99.5% identity in 186 amino acid overlap with the carboxyl terminus of calpastatin. The BS-17 component of human sperm corresponds to the carboxyl terminus of calpastatin. This conclusion is supported by the finding that the polyclonal antibodies also interacted with a 84 kD protein corresponding to the M(r) of calpastatin.
从一名不孕女性血清中提取的抗体,通过蛋白质免疫印迹法与从人类精子中提取的一种17.5 kD糖蛋白(BS - 17成分)发生相互作用。制备了针对BS - 17成分的多克隆抗体,并用于从人类睾丸λgt11表达文库中鉴定阳性染色克隆。分离出编码BS - 17成分的cDNA并测定其核苷酸序列。BS - 17 cDNA包含758个核苷酸,其开放阅读框为558个核苷酸,编码一个由186个氨基酸组成的多肽。在758个核苷酸的重叠区域,BS - 17 cDNA与编码钙蛋白酶抑制蛋白的基因3'末端显示出99.7%的同源性,在186个氨基酸的重叠区域与钙蛋白酶抑制蛋白的羧基末端具有99.5%的同一性。人类精子的BS - 17成分对应于钙蛋白酶抑制蛋白的羧基末端。这一结论得到以下发现的支持:多克隆抗体还与一种对应于钙蛋白酶抑制蛋白分子量的84 kD蛋白质发生相互作用。