Outer membrane proteins of Bartonella henselae and their interaction with human endothelial cells.
Members of the genus Bartonella are unique in that they are bacteria which cause proliferation of microvascular endothelial cells and neovascularization (angiogenesis). The mechanisms by which Bartonella henselae causes these processes are unknown. Given the importance of surface-exposed determinants in the pathogenesis of many organisms, outer membrane proteins (OMPs) of B. henselae were identified. Enrichment of the outer membrane fraction of B. henselae by sarkosyl treatment of total membranes, together with radioiodination and biotinylation of intact organisms, suggest that at least nine proteins, with molecular weights of 28, 30, 35, 43, 58, 61, 79, 92 and 171 kDa, are located in the outer membrane. Triton X-100-extracted biotinylated human umbilical vein endothelial cell (HUVEC) surface proteins bound to the 43 kDa B. henselae OMP after B. henselae whole-cell lysates and sarkosyl-fractionated OMPs were separated by SDS-PAGE and transferred onto nylon. Biotinylated B. henselae surface proteins of 28, 32, 43, 52 and 58 kDa were shown to bind intact HUVEC, with the 43 kDa protein being the major adhesin. Preincubation of HUVEC with an increasing concentration (20 microg/ml to 4 mg/ml) of sarkosyl-fractionated unlabelled B. henselae outer membrane proteins inhibited the attachment of all identified HUVEC binding proteins. The identification of B. henselae OMPs, as well as adhesins, should provide a basis for further investigation of the role of adherence in the pathogenesis of B. henselae.
Burgess
A W
AW
University of South Florida College of Medicine, Department of Medical Microbiology and Immunology, 12901 Bruce B. Downs Boulevard, Tampa, Florida, 33612, USA.
Anderson
B E
BE
D018829:Adhesins, Bacterial / Q000737:chemistry; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D018416:Bartonella henselae / Q000737:chemistry / Q000254:growth & development / Q000472:pathogenicity*; D019921:Biotinylation; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000382:microbiology*; D006801:Humans; D014471:Umbilical Veins / Q000166:cytology
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins