Zhang Wen-Ming, Wang Hai-Feng, Gao Kan, Wang Cong, Liu Li, Liu Jian-Xin
College of Animal Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou 310029, People's Republic of China.
Can J Microbiol. 2015 May;61(5):373-80. doi: 10.1139/cjm-2014-0734. Epub 2015 Mar 23.
This study was aimed to identify key surface proteins mediating the adhesion of lactobacilli to intestinal epithelial cells. By using Caco-2 and IPEC-J2 cells labeled with sulfo-NHS-biotin in the western blotting, a protein band of an approximately 37 kDa was detected on the surface layer of Lactobacillus reuteri strains ZJ616, ZJ617, ZJ621, and ZJ623 and Lactobacillus rhamnosus GG. Mass spectrometry analysis using the adhesion-related protein from L. reuteri ZJ617 showed that it was 100% homologous to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of L. reuteri JCM 1112 (GenBank: YP_001841377). The ability of L. reuteri ZJ617 to adhere to epithelial cells decreased significantly by treatment with LiCl or by blocking with an anti-GAPDH antibody, in comparison with the untreated strain (p < 0.05). Immunoelectron microscopic and immunofluorescence analyses confirmed that GAPDH is located on the surface layer of L. reuteri ZJ617. The results indicated that the GAPDH protein of L. reuteri ZJ617 acts as an adhesion component that plays an important role in binding to the intestinal epithelial cells.
本研究旨在鉴定介导乳酸菌与肠上皮细胞黏附的关键表面蛋白。通过在蛋白质印迹法中使用经磺基-NHS-生物素标记的Caco-2和IPEC-J2细胞，在罗伊氏乳杆菌菌株ZJ616、ZJ617、ZJ621和ZJ623以及鼠李糖乳杆菌GG的表面层检测到一条约37 kDa的蛋白条带。对来自罗伊氏乳杆菌ZJ617的黏附相关蛋白进行质谱分析表明，它与罗伊氏乳杆菌JCM 1112（GenBank：YP_001841377）的甘油醛-3-磷酸脱氢酶（GAPDH）100%同源。与未处理的菌株相比，用LiCl处理或用抗GAPDH抗体封闭后，罗伊氏乳杆菌ZJ617黏附上皮细胞的能力显著降低（p < 0.05）。免疫电子显微镜和免疫荧光分析证实GAPDH位于罗伊氏乳杆菌ZJ617的表面层。结果表明，罗伊氏乳杆菌ZJ617的GAPDH蛋白作为一种黏附成分，在与肠上皮细胞结合中起重要作用。
