Tkalcevic J, Ashman K, Meeusen E
Centre for Animal Biotechnology, University of Melbourne, School of Veterinary Science, Parkville, Victoria, Australia.
Biochem Biophys Res Commun. 1995 Aug 4;213(1):169-74. doi: 10.1006/bbrc.1995.2112.
The sensitivity of N-terminal sequencing has been used to identify proteins expressed by the newly excysted juvenile stage of the parasite Fasciola hepatica. Of the seven proteins identified, a number have significant sequence homology to the cysteine proteases: cathepsin B, cathepsin L and asparaginyl endoproteinase. Proteolytic activity was demonstrated using gelatin substrate sodium dodecyl sulphate polyacrylamide gel electrophoresis. In addition, a number of novel proteins were identified which shared no significant sequence homology to proteins in the databases. The availability of such N-terminal sequence information allows rapid identification of major proteins from scarce developmental stages and provides the basis for further molecular studies.
N端测序的敏感性已被用于鉴定肝片吸虫寄生虫新脱囊幼虫阶段表达的蛋白质。在鉴定出的7种蛋白质中,有几种与半胱氨酸蛋白酶具有显著的序列同源性:组织蛋白酶B、组织蛋白酶L和天冬酰胺基内肽酶。使用明胶底物十二烷基硫酸钠聚丙烯酰胺凝胶电泳证明了蛋白水解活性。此外,还鉴定出了一些与数据库中的蛋白质没有显著序列同源性的新蛋白质。此类N端序列信息的可用性使得能够从稀缺的发育阶段快速鉴定主要蛋白质,并为进一步的分子研究提供基础。