Synthesis and secretion of proteins by perifused caput epididymal tubules, and association of secreted proteins with spermatozoa.
Synthesis and secretion of proteins by perifused caput epididymal tubules, and association of secreted proteins with spermatozoa.
复制标题
灌注头附睾小管蛋白质的合成和分泌，以及分泌蛋白质与精子的关联。
DOI：
10.1095/biolreprod33.4.1017
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发表时间：
1985
影响因子：
3.6
通讯作者：
Hamilton,DW
中科院分区：
文献类型：
--
作者：
Klinefelter,GR;Hamilton,DW
本实验采用大鼠附睾头近端和远端灌流器官培养法，研究了附睾上皮蛋白质的分泌和精子对管腔内放射性蛋白质的摄取。L-[35 S]蛋氨酸掺入管腔液蛋白的量是时间依赖性的，并完全抑制放线菌酮。标记蛋白与培养精子的结合也依赖于时间和连续性，在蛋白合成被抑制后，精子仍然获得标记的管腔蛋白。在管腔液和精子提取物中发现了一个Mr= 46，000的分子被大量标记。使用抗附睾α-乳白蛋白抗体免疫沉淀的所有L-[35 S]蛋氨酸提取物的荧光图（Klinefelter和汉密尔顿，1984）显示标记了Mr= 18，000的分子，此外还标记了Mr= 46，000的分子，但免疫染色仅对Mr= 18，000的分子和免疫球蛋白重链具有特异性。用半乳糖氧化酶-NaB [~ 3 H]_4标记培养的精子头细胞表面，发现一个Mr= 23，000的分子能与抗附睾α-乳白蛋白抗体发生免疫沉淀。我们的数据表明，这种细胞表面分子是类似的流体附睾α-乳白蛋白样复合物的一个组成部分，此外，表明精子表面的糖基化可以发生在附睾头。
We have used perifusion organ culture of proximal and distal caput epididymal tubules of the rat to study the secretion of proteins by epididymal epithelium and uptake of the luminal radioactive proteins by sperm. The amount of incorporation of L-[35S]methionine into luminal fluid proteins was time dependent and completely inhibited by cycloheximide. The association of labeled proteins with cultured sperm was also dependent on time and continuous, with sperm still acquiring labeled luminal proteins after protein synthesis was arrested.A Mr= 46,000 molecule was found to be heavily labeled in luminal fluid and sperm extracts. Fluorograms of all L-[35S]methionine extracts immunoprecipitated using an antiepididymal alpha-lactalbumin antibody (Klinefelter and Hamilton, 1984) showed labeling of an Mr= 18,000 molecule and, in addition, the Mr= 46,000 molecule, but immunostaining was specific only for the Mr= 18,000 molecule and the heavy chain of the immunoglobulin. We suggest that the Mr= 46,000 molecule may be galactosyltransferase.Galactose oxidase-NaB[3H]4labeling of the cultured caput sperm cell surface revealed a Mr= 23,000 molecule that was able to be immunoprecipitated with antiepididymal alpha-lactalbumin antibody. Our data suggest that this cell surface molecule is similar to one component of the fluid epididymal alpha-lactalbumin-like complex and, in addition, show that glycosylation of the sperm surface can occur in the caput epididymidis.