title: Whole surface image of Mycoplasma mobile, suggested by protein identification and immunofluorescence microscopy.
abstract: Mycoplasma mobile, a freshwater fish pathogen featured with robust gliding motility, binds to the surface of the gill, where it then colonizes. Here, to obtain a whole image of its cell surface, we identified the proteins exposed on the surface using the following methods. (i) The cell surface was labeled with sulfosuccinimidyl-6-(biotinamido) hexanoate and recovered by an avidin column. (ii) The cells were subjected to phase partitioning using Triton X-114, and the hydrophobic proteins were recovered. (iii) The membrane fraction was analyzed by two-dimensional gel electrophoresis. These recovered proteins were subjected to peptide mass fingerprinting, and a final list of 36 expressed surface proteins was established. The ratio of identified proteins to whole surface proteins was estimated through two-dimensional gel electrophoresis of the membrane fraction. The localization of three newly found proteins, Mvsps C, E, and F, has been clarified by immunofluorescence microscopy. Integrating all information, a whole image of the cell surface showed that the proteins for gliding that were localized at the base of the protrusion of flask-shaped M. mobile account for more than 12% of all surface proteins and that Mvsps, surface variants that were localized at both parts other than the neck, account for 49% of all surface proteins.
authors.affiliation: Department of Biology, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka, Japan.
authors.forename: Heng Ning
authors.initials: HN
authors.lastname: Wu
authors.forename: Makoto
authors.initials: M
authors.lastname: Miyata
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis*; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D008856:Microscopy, Fluorescence; D009174:Mycoplasma / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins