Sarkar M, Majumder G C, Chatterjee T
Indian Institute of Chemical Biology, Calcutta.
Biochim Biophys Acta. 1991 Nov 18;1070(1):198-204. doi: 10.1016/0005-2736(91)90164-4.
The cell surface glycoproteins of goat epididymal maturing spermatozoa have been investigated using lectins as surface probes that interact with specific sugars with high affinity. Concanavalin A (ConA) and wheat-germ agglutinin (WGA) showed high affinity for mature cauda epididymal sperm agglutination, whereas RCA2, kidney beans lectin and peanut agglutinin caused much lower or little agglutination of the cells. The mature sperm exhibited markedly higher efficacy than the immature caput epididymal sperm for binding both ConA and WGA, as evidenced by sperm agglutination and the binding of the fluorescence isothiocyanate (FITC)-labelled lectins. FITC-ConA binds uniformly to the entire mature sperm surface whereas FITC-WGA binds to the acrosomal cap region of the head. The FITC-RCA2 mainly labelled the posterior head of mature cauda sperm. However, no WGA-specific glycoprotein receptors could be detected in sperm plasma membrane (PM) by WGA-Sepharose affinity chromatography. The data implied that the epididymal sperm maturation is associated with a marked increase in the ConA/WGA receptors and that WGA receptors may be glycolipids rather than glycoproteins. Analysis of the ConA receptors of cauda sperm PM identified by ConA-Sepharose affinity chromatography and subsequent resolution in SDS-PAGE demonstrated the presence of five glycopolypeptides of different concentrations (98, 96, 43, 27 and 17 kDa) of goat sperm membrane. The immunoblot of these ConA-specific glycopeptides with anti-sperm membrane antiserum showed that 98- and 96-kDa receptors are immunoresponsive.
利用凝集素作为与特定糖类具有高亲和力的表面探针，对山羊附睾成熟精子的细胞表面糖蛋白进行了研究。刀豆球蛋白A（ConA）和麦胚凝集素（WGA）对成熟附睾尾精子的凝集表现出高亲和力，而蓖麻凝集素2（RCA2）、菜豆凝集素和花生凝集素对细胞的凝集作用则低得多或几乎没有。成熟精子对ConA和WGA的结合能力明显高于未成熟的附睾头精子，精子凝集以及异硫氰酸荧光素（FITC）标记的凝集素的结合证明了这一点。FITC-ConA均匀地结合在整个成熟精子表面，而FITC-WGA则结合在头部的顶体帽区域。FITC-RCA2主要标记成熟附睾尾精子的头部后部。然而，通过WGA-琼脂糖亲和层析在精子质膜（PM）中未检测到WGA特异性糖蛋白受体。数据表明，附睾精子成熟与ConA/WGA受体的显著增加有关，并且WGA受体可能是糖脂而不是糖蛋白。通过ConA-琼脂糖亲和层析鉴定并随后在SDS-PAGE中分离的附睾尾精子PM的ConA受体分析表明，山羊精子膜存在五种不同浓度（98、96、43、27和17 kDa）的糖多肽。用抗精子膜抗血清对这些ConA特异性糖肽进行免疫印迹分析表明，98 kDa和96 kDa的受体具有免疫反应性。
