摘要
目的初步掌握流行于青海省细粒棘球绦虫幼虫棘球蚴(原头节、囊壁)在中间宿主人体内蛋白质表达情况。方法利用SDS-PAGE和western-blot分析棘球蚴原头节和囊壁蛋白质表达谱,进一步利用2DE和MALDI-TOF质谱分析技术对原头节和囊壁蛋白质构成情况进行全面的研究分析。结果在原头蚴、囊壁组织的双向电泳图中,各蛋白点通过MALDI-TOF-MS检测,原头蚴和囊壁分别有40个蛋白质点和8个蛋白质点得以初步鉴定,包括有:(1)细胞骨架蛋白:肌动蛋白、原肌球蛋白。(2)代谢相关酶类:苹果酸脱氢酶;磷酸丙糖异构酶;脯氨酸氧化酶;谷胱甘肽-S-转移酶;类胡萝卜脱氢酶;肽基脯氨酰顺反异构酶;(3)物质转运相关蛋白:载脂蛋白A-I;(4)应激反应蛋白(HSP70,HSP20相关蛋白)。结论初步鉴定了寄生人体细粒棘球蚴原头蚴、囊壁组织中的部分蛋白质点,其中,原头蚴中共有40个蛋白点,囊壁中有8个蛋白点。这些蛋白可能与细粒棘球蚴运动、生长、发育、代谢调控等方面有关。
The Qinghai Tibetan Plateau in western China, which is the world's largest and highest plateau, is a highly epidemic area for Echinococcus spp. To investigate the profiles at the protein expression level, proteomics analysis were carried out by one dimension and two dimension polyacrylamide gel electrophoresis to detect the protein expressing profiles of proto- scoleees and cyst wall. All protein spots were visualized by using Coomassie Blue G-250. Reproducible abundant protein spots in three parallel gels were detected on each 2DE gels with PI (3-10) by PDQuest software. Of these proteins, 40 of proto- scoleees and 8 of cyst wall were analyzed by mass spectrometry and identified in the NCBInr database. The foundational categories of identified proteins were: (1) hydatid disease diagnostic antigen P29; (2) 14-3-3 proteim (3) carbohydrate/energy metabolism; (4) cell membrane/cytoskeleton. The proteomics analysis of protoscoleces and cyst wall of Echinococcus granulosus could bring us into a new insight in our exploring on the biological properties, immunogenicity and metabolism characteristics of Echinococcus granulosus endemic in Qinghai Tibetan Plateau.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2014年第6期575-577,582,共4页
Chinese Journal of Zoonoses
基金
国家自然科学基金地区项目(No.81360255)
春晖计划项目(No.2012-Z082)
青海大学中青年基金项目(No.2012-AYY-4)
青海大学医学院中青年科研基金项目(No.qykj-01)联合资助~~
关键词
细粒棘球绦虫
棘球蚴原头节
囊壁
蛋白质表达谱
Echinococcus granulosus
hydatid disease
protoscoleces
cyst wall
protein expressing profiles