小白鼠貯精囊自體抗原的特性研究

Abstract

中文摘要 本實驗室從前發現純系（Balb / c）小白鼠的貯精囊中，存在－個蛋白質，將其注射到雄性或雌性的小白鼠，都可以引起免疫反應，因此命名為貯精囊自體抗原（Seminal Vesicles Auto antigen , SVA ）。它是一個醣蛋白，分子量由 SDS － PAGE 估計為 30K ，由 gelfiltration 估計為 22k ，其 N 端前 20 個氨基酸序列與已知的蛋白質比對，並無任何明顯的相似性，猜測為一未被研究的蛋白質。本實驗繼續 SVA 的特性研究，首先藉由兔子製備 Anti－SVA 抗體，以供免疫分析使用。其次對 SVA 的醣蛋白本質做進一步的研究，瞭解 SVA 的醣基佔重量的 20 . 6 ％，可能含有兩個或兩個以上的 N－1inked oligosaccharides 及若干 O－linked oligosaccharides 。此外利用胰蛋白?水解 SVA ，得到若干Pept ides ，並得到其中兩段的氨基酸序列，並且與己完成的 cDNA 序列相吻合。 最後利用未成熟的幼鼠及閹割的成鼠，證明 SVA 受到雄性素的調控。

Summary The previous investigation of our group had resulted in purifying an auto antigen from the semifinal vesicles of inbred mice (Balb/c). We named it Seminal Vesicles Auto antigen (SVA). It was a glycoprotein and its molecular weight was estimated about 30k according to SDS-PAGE or 22k according to gel filtration. Based on the sequence of 20 amino acid in the N terminal region, SVA shows no significant homology with know protein sequences. So it could be considered as a new protein. In this work, we characterized SVA further. The Rabbitanti-SVA were prod used. The production of SVA was shown to be androgen-dependent by immunochemical detection. The glycoprotein nature of SVA was studied further. We found that the oligosaccharide moiety of SVA constituted about 20.6% of total SVA mass. Both N-linked and 0-linked oligosaccharides were found in SVA molecule. Two or more than two sites on SVA molecule might be involved in the congugation of the N-linked oligosaccharide to the protein core that has molecular weight of 15.3k. The trypsin-digested peptides of the reductive-alkylated SVA were resolved by HPLC. Two of them gave the amino acid sequences confirmed the one deduced from cDNA sequence.