SCHISTOSOMA-MANSONI - CHARACTERIZATION OF EXCRETORY-SECRETORY POLYPEPTIDES SYNTHESIZED IN-VITRO BY DAUGHTER SPOROCYSTS
SCHISTOSOMA-MANSONI - CHARACTERIZATION OF EXCRETORY-SECRETORY POLYPEPTIDES SYNTHESIZED IN-VITRO BY DAUGHTER SPOROCYSTS
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DOI：
10.1006/expr.1995.1004
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发表时间：
1995-02-01
影响因子：
2.1
通讯作者：
YOSHINO, TP
中科院分区：
文献类型：
--
作者：
CREWSOYEN, AE;YOSHINO, TP
子孢子囊的排泄分泌（ES）产物参与蜗牛宿主生殖生理的调节。由于ES产物在蜗牛繁殖中的潜在重要作用，我们使用[S-35]甲硫氨酸进行脉冲追踪研究，然后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、荧光照相和扫描密度测定，检查了体外培养的曼氏血吸虫子孢子囊新合成的ES多肽。SDS-PAGE分离的ES蛋白的荧光图显示，孢子囊在26 ℃下体外培养6天期间向上清液中释放分子量范围为17至200 kDa的多种多肽。此外，子孢子囊多肽合成和释放到培养上清液中的一般模式不同于先前描述的来自初级或母孢子囊的ES组分。虽然定量光密度分析显示，总标记ES蛋白释放到培养上清液中是一致的三个脉冲追逐期间，有个别多肽峰的数量显着差异。选择用于进一步分析的八种多肽的合成和释放的动力学随时间变化。几个ES多肽（110，95，和25 kDa）被释放在相对恒定的量，而一些（69和44 kDa）增加和其他（38和35 kDa）在同一时期的文化减少的数量。据推测，38-和35-kDa的多肽可能是重要的候选抑制分子，因为它们的早期释放到培养上清液和第1-2天的子孢子囊ES产物对蜗牛宿主的蛋白腺中的多糖合成的抑制作用之间的相关性。该研究确定了亲核体子孢子囊能够调节某些多肽的合成和释放到培养上清液中。次级孢子囊ES产物在改变宿主-寄生虫生理相互作用的各个方面中的可能作用被讨论，（C）1995 Academic Press，Inc.
Excretory-secretory (ES) products of daughter sporocysts have been implicated in the modulation of snail host reproductive physiology. Because of the potentially important role of ES products in snail reproduction, newly synthesized ES polypeptides of in vitro-cultured Schistosoma mansoni daughter sporocysts were examined in pulse-chase studies using [S-35]methionine followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), fluorography, and scanning densitometry. Fluorograms of SDS-PAGE-separated ES proteins revealed that sporocysts released a wide variety of polypeptides ranging in molecular weight from 17 to 200 kDa into supernatants during 6 days of in vitro culture at 26 degrees C. Moreover, the general pattern of synthesis and release of daughter sporocyst polypeptides into culture supernatants differed from those ES components previously described from primary or mother sporocysts. Although quantitative densitometry revealed that total labeled ES protein released into culture supernatants was consistent for an three pulse-chase periods, there were significant differences in the quantities of individual polypeptide peaks. The dynamics of the synthesis and release of eight polypeptides chosen for further analysis varied over time. Several ES polypeptides (110, 95, and 25 kDa) were released in relatively constant amounts, while some (69 and 44 kDa) increased and others (38 and 35 kDa) decreased in quantity over the same period of culture. It is hypothesized that the 38- and 35-kDa polypeptides may be important candidate inhibitory molecules because of the correlation between their early release into culture supernatants and the inhibitory effect of Day 1-2 daughter sporocyst ES products on polysaccharide synthesis in the albumen gland of the snail host. This study firmly establishes that schistosome daughter sporocysts are able to regulate the synthesis and release of certain polypeptides into culture supernatants. The possible roles of secondary sporocyst ES products in altering various aspects of host-parasite physiological interactions are discussed, (C) 1995 Academic Press, Inc.