口腔副溶血链球菌黏附蛋白糖基化相关基因产物的亚细胞定位及黏附功能研究  

Subcellular localization and in vitro adhesion experiment of Streptococcus parasanguinis adhesin glycosylation-associated gene mutants

作　　者：彭志翔[1] Hui Wu Paula Fives-Taylor Baiming Sun 

机构地区：[1]中山大学附属口腔医院牙体牙髓科,广州510055 [2]美国亚拉巴马大学牙医学院及医学院,伯明翰VT05405 [3]美国佛蒙特大学微生物及分子遗传学系,伯灵顿AL35294

出　　处：《口腔生物医学》2010年第2期61-64,共4页Oral Biomedicine

基　　金：美国NIH赞助基金(DE11000;DE017954)

摘　　要：目的调查口腔副溶血链球菌黏附蛋白Fap1糖基化相关基因产物Gap1、Gap2和Gap3的亚细胞定位,以及相关基因缺陷株的黏附能力改变情况。方法等位置换技术获得口腔副溶血链球菌黏附蛋白糖基化相关基因缺陷株gap1-、gap2-和gap3-,穿梭质粒构建该三种基因的补偿株,Western Blot检测Gap1、Gap2和Gap3在副溶血链球菌内的亚细胞定位;闪烁计数法检测gap1-、gap2-和gap3-对羟基磷灰石的黏附能力。结果 Gap1和Gap2被发现分布于所有亚细胞组分中,但主要集中于胞浆和胞膜内,而Gap3仅分布于胞浆和胞膜;体外黏附实验显示gap1-、gap2-和gap3-对羟基磷灰石的黏附能力显著下降。结论糖基化修饰对副溶血链球菌黏附蛋白Fap1的黏附功能至关重要;Gap1、Gap2和Gap3可能在Fap1糖基化修饰过程中协同作用,该协同作用发生在胞内环境。Objective To investigate the subcellular localization of Streptococcus parasanguinis adhesin Fapl glycosylation-associated gene products,and the adhesion capacity of the glycosylation-associated gene mutants. Methods Insertional replacement technique was used to construct Fapl glycosylation-associated gene defect mutants gap1 - ,gap2 - and gap3 -. Their complementary strains were constructed with a shuttle plasmid. Western Blot revealed the suhcellular localization of Gap1, Gap2 and Gap3 in Streptococcus parasanguinis. Scintillation Counting was carried out to access the adhesion capacity of all mutants. Results Gapl and Gap2 were detected in all subcellular fractions but were distributed predominately in cytoplasm and membrane, whereas Gap3 was found only in cytoplasm and membrane. The in vitro adhesion data indicated that these 3 mutants had obviously decreased binding activities to SHA. Conclusions Glycosylation modification on adhesin Fapl of Streptococcus parasanguinis is important for its adhesion capacity. Gap proteins may function cooperatively during Fap1 glycosylation process. Such a joint function may take place in an intracellular environment.

关 键 词：副溶血链球菌 糖基化相关基因 亚细胞定位 黏附功能 

分 类 号：R780.2[医药卫生—口腔医学]