Isolation and Characterization of the Outer Membrane of Borrelia hermsii

摘要: The outer membrane of Borrelia hermsii has been shown by freeze-fracture analysis to contain a low density membrane-spanning proteins which have not yet isolated or identified. In this study, we report the purification vesicles (OMV) from B. HS-1 and subsequent identification their constituent proteins. membranes were released vigorous vortexing whole organisms in low-pH, hypotonic citrate buffer isopycnic sucrose gradient centrifugation. OMV exhibited porin activities ranging 0.2 7.2 nS, consistent with origin. Purified be relatively free inner contamination absence measurable β-NADH oxidase activity protoplasmic cylinder-associated observed Coomassie blue staining. Approximately 60 protein spots (some are putative isoelectric isomers) 25 distinct molecular weights identified as constituents enrichment. majority these also antigenic sera -infected mice. Seven labeled [ 3 H]palmitate, including surface-exposed glycerophosphodiester phosphodiesterase, variable major 7 33, 15, 17, 38, 42, 67 kDa, indicating that they lipoprotein membrane. addition, immunoblot probed antiserum garinii p66/Oms66 demonstrated presence p66 (Oms66) homolog. Treatment intact proteinase K resulted partial proteolysis Oms66/p66 homolog, it is surface exposed. This characterization should aid further studies pathogenesis immunity tick-borne relapsing fever.