Adhesions of extracellular surface-layer associated proteins in Lactobacillus M5-L and Q8-L
Autor: | Linlin Wang, Yingchun Zhang, Qianhui Lu, Xinling Xiang, Lanwei Zhang, Fang Ma |
---|---|
Rok vydání: | 2015 |
Předmět: |
0301 basic medicine
Lactobacillus casei Lactobacillus paracasei Blotting Western Biotin Succinimides Biology Bacterial Adhesion 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins Lactobacillus Genetics Extracellular Animals Humans Membrane Glycoproteins Strain (chemistry) food and beverages Adhesion Gene Expression Regulation Bacterial biology.organism_classification Blot Intestines 030104 developmental biology Biochemistry chemistry Animal Science and Zoology Electrophoresis Polyacrylamide Gel HT29 Cells Food Science |
Zdroj: | Journal of dairy science. 99(2) |
ISSN: | 1525-3198 |
Popis: | Surface-layer associated proteins (SLAP) that envelop Lactobacillus paracasei ssp. paracasei M5-L and Lactobacillus casei Q8-L cell surfaces are involved in the adherence of these strain to the human intestinal cell line HT-29. To further elucidate some of the properties of these proteins, we assessed the yields and expressions of SLAP under different incubation conditions. An efficient and selective extraction of SLAP was obtained when cells of Lactobacillus were treated with 5 M LiCl at 37°C in aerobic conditions. The SLAP of Lactobacillus M5-L and Q8-L in cell extracts were visualized by SDS-PAGE and identified by Western blotting with sulfo-N-hydroxysuccinimide-biotin-labeled HT-29 cells as adhesion proteins. Atomic force microscopy contact imaging revealed that Lactobacillus strains M5-L and Q8-L normally display a smooth, homogeneous surface, whereas the surfaces of M5-L and Q8-L treated with 5 M LiCl were rough and more heterogeneous. Analysis of adhesion forces revealed that the initial adhesion forces of 1.41 and 1.28 nN obtained for normal Lactobacillus M5-L and Q8-L strains, respectively, decreased to 0.70 and 0.48 nN, respectively, following 5 M LiCl treatment. Finally, the dominant 45-kDa protein bands of Lactobacillus Q8-L and Lactobacillus M5-L were identified as elongation factor Tu and surface antigen, respectively, by liquid chromatography-tandem mass spectrometry. |
Databáze: | OpenAIRE |
Externí odkaz: |