Synthesis and processing of rat sperm-associated alpha-L-fucosidase.
Synthesis and processing of rat sperm-associated alpha-L-fucosidase.
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大鼠精子相关α-L-岩藻糖苷酶的合成和加工。
DOI：
10.1095/biolreprod48.6.1228
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发表时间：
1993
影响因子：
3.6
通讯作者：
AronsonJr,NN
中科院分区：
文献类型：
--
作者：
Hancock,LW;Raab,LS;AronsonJr,NN
α-L-岩藻糖苷酶（α-岩藻糖苷酶）是通过使用人工4-甲基伞形酮基底物在大鼠附睾精子中测定的多种糖苷酶中最具活性的。此外，在纯化的睾丸生殖细胞群中可检测到酶活性;与圆形精子细胞相比，附睾头精子每个细胞的α-岩藻糖苷酶活性至少增加1.8倍。对培养的睾丸生殖细胞进行代谢标记，然后进行免疫沉淀和SDS-PAGE，结果显示粗线期精母细胞和圆形精子细胞中合成了54 kDa多肽α-岩藻糖苷酶，但在凝聚精子细胞中没有合成。该多肽是N-糖基化的，但不经历进一步加工，如通过脉冲/追踪标记和用内切糖苷酶H处理所确定的。相比之下，由培养的克隆9细胞（大鼠肝源性细胞系）合成的α-岩藻糖苷酶经历了从54 kDa前体到SDS-PAGE上以56 kDa为中心的稍大中间体（通过碳水化合物修饰）的加工，最后加工成52 kDa成熟多肽。免疫印迹证实了睾丸生殖细胞中存在54 kDa形式的α-岩藻糖苷酶，并显示附睾精子中存在52 kDa成熟多肽。此外，免疫反应性多肽在附睾头精子制备物中更突出;这与酶活性的增加一致。在圆形精子细胞发育阶段之后缺乏α-岩藻糖苷酶合成的情况下，似乎α-岩藻糖苷酶可能是精子在通过流出管系统的过程中从腔液中获得的。代谢标记的培养附睾上皮细胞合成并分泌免疫可沉淀的58-kDa α-岩藻糖苷酶多肽到培养基中的证据支持了这一假设。对从附睾头第一部分分离的精子进行的分析表明，精子相关α-岩藻糖苷酶的获得和加工发生在精子到达附睾之前或同时。
α-L-Fucosidase (α-fucosidase) is the most active of a number of glycosidases measured in rat epididymal sperm through use of artificial 4-methylumbelliferyl substrates. In addition, enzyme activity can be detected in purified populations of testicular germ cells; in comparison to round spermatids, caput epididymal sperm show at least a 1.8-fold increase in α-fucosidase activity per cell. Metabolic labeling of cultured testicular germ cells, followed by immunoprecipitation and SDS-PAGE, reveals synthesis of α-fucosidase as a polypeptide of 54 kDa in pachytene spermatocytes and round spermatids but no synthesis in condensing spermatids. This polypeptide is N-glycosylated but does not undergo further processing as determined by pulse/chase labeling and treatment with endoglycosidase H. In contrast, the α-fucosidase synthesized by cultured clone 9 cells (a rat liver-derived cell line) undergoes processing from a 54-kDa precursor to a slightly larger intermediate centered at 56 kDa on SDS-PAGE (via carbohydrate modification) and finally to a 52-kDa mature polypeptide. Immunoblotting confirms the presence of the 54-kDa form of α-fucosidase in testicular germ cells and shows the existence of a 52-kDa mature polypeptide in epididymal sperm. In addition, immunoreactive polypeptide is more prominent in caput epididymal sperm preparations; this is consistent with the increase in enzyme activity. In the absence of α-fucosidase synthesis beyond the round spermatid stage of development, it appears that α-fucosidase may be acquired from the luminal fluid by sperm during transit through the excurrent duct system. This hypothesis is supported by evidence that metabolically labeled cultured epididymal epithelial cells synthesize and secrete into culture medium an immunoprecipitable 58-kDa α-fucosidase polypeptide. Analysis of sperm isolated from the first part of the caput epididymis indicates that both acquisition and processing of sperm-associated α-fucosidase takes place prior to or concomitant with arrival of sperm in the epididymis.
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岩藻多糖通过与内顶体膜和赤道区结合，抑制豚鼠精子附着在透明带上。
影响因子：
3.7
作者：
Huang,TT;Yanagimachi,R
通讯作者：
Yanagimachi,R
影响因子：
3.6
作者：
SYLVESTER, SR;MORALES, C;GRISWOLD, MD
通讯作者：
GRISWOLD, MD
在 Celsep (TM) 装置上分离小鼠睾丸细胞及其作为高分子量 DNA 或 RNA 来源的用途。
DOI：
10.1002/mrd.1120120102
发表时间：
1985
期刊：
Gamete research
影响因子：
--
作者：
Wolgemuth,DJ;Gizang-Ginsberg,E;Engelmyer,E;Gavin,BJ;Ponzetto,C
通讯作者：
Ponzetto,C
影响因子：
--
作者：
Luis S. Mayorga;Francisco Bertini
通讯作者：
Francisco Bertini
与附睾运输相关的大鼠精子膜糖苷酶活性以及碳水化合物和蛋白质含量的变化。
影响因子：
3.6
作者：
Joseph C. Hall;Gary J. Killian
通讯作者：
Gary J. Killian