Nikolajczyk B S, O'Rand M G
Department of Cell Biology and Anatomy, University of North Carolina, Chapel Hill 27599.
Biol Reprod. 1992 Mar;46(3):366-78. doi: 10.1095/biolreprod46.3.366.
Examination of the role of carbohydrates in specific recognition between spermatozoa and zona pellucida has focussed on understanding the interaction of sperm hydrolases or lectin-like molecules with zona pellucida ligands. To elucidate the role of specific spermatozoan hydrolases in gamete interaction, rabbit testis beta-galactosidase and arylsulfatase A were purified, characterized, and localized in spermatozoa. beta-Galactosidase and arylsulfatase A co-purified after affinity, size, or reverse-phase chromatography. N-Terminal amino acid analysis and enzymatic characterization suggested that neither enzyme is a testis-specific isozyme. Size chromatography indicated that both enzymes aggregated into macromolecular complexes at pH 4.0, while both dissociated at pH 8.0. beta-Galactosidase and arylsulfatase A co-localized on the sperm surface and in the acrosome and postacrosomal regions of spermatozoa. Throughout the zona-induced acrosome reaction, both enzymes remained associated with the detached acrosomal cap and postacrosomal region of acrosome-reacted spermatozoa. Because the acrosome is an acidic subcellular compartment, internal beta-galactosidase and arylsulfatase A are probably aggregated in acrosome-intact spermatozoa and dissociate as they are exposed to pH increases during the acrosome reaction.
对碳水化合物在精子与透明带之间特异性识别中所起作用的研究，聚焦于了解精子水解酶或类凝集素分子与透明带配体之间的相互作用。为阐明特定精子水解酶在配子相互作用中的作用，对兔睾丸β-半乳糖苷酶和芳基硫酸酯酶A进行了纯化、特性鉴定并定位在精子中。β-半乳糖苷酶和芳基硫酸酯酶A在亲和色谱、尺寸排阻色谱或反相色谱后共同纯化。N端氨基酸分析和酶学特性表明这两种酶都不是睾丸特异性同工酶。尺寸排阻色谱表明，两种酶在pH 4.0时聚集成大分子复合物，而在pH 8.0时都解离。β-半乳糖苷酶和芳基硫酸酯酶A共同定位于精子表面以及精子的顶体和顶体后区域。在整个透明带诱导的顶体反应过程中，这两种酶都与顶体反应精子分离的顶体帽和顶体后区域保持关联。由于顶体是一个酸性亚细胞区室，内部的β-半乳糖苷酶和芳基硫酸酯酶A在顶体完整的精子中可能聚集，而在顶体反应过程中随着pH升高而暴露时会解离。
