The ability of bacteria to adhere to and colonize host cells is mediated by specific interactions between bacterial surface components (adhesins) and complementary host cell surface receptors including the extracellular matrix (ECM) molecules fibronectin (Fn) and heparin sulfate (HS). Fn is a glycoprotein that binds several ECM molecules and is found in soluble form in plasma and in fibrillar form in the ECM and basement membranes. Glycosaminoglycans (GAGs) such as heparin (Hep) and HS can also serve as putative receptors (1, 49). Hep and HS have common structural features and participate in the GAG-mediated recruitment of host ECM protein, a novel strategy in bacterial adherence and invasion demonstrated by Duensing et al. (15). HS, the most ubiquitous member of the GAGs, is found on the surfaces of most eukaryotic cells as part of the ECM, whereas Hep is synthesized in mast cells and basophils.For several pathogens, the binding to ECM molecules such as Fn and Hep is an important step in pathogenesis that involves specific bacterial surface proteins (16,26,33,47). This is particularly true for wound-associated infections such as those caused by Bartonella henselae. B. henselae, an opportunistic and emerging pathogen, is carried by cats and causes several distinct clinical syndromes in immunocompetent individuals and AIDS patients including cat scratch disease, bacillary angiomatosis, chronic bacteremia, and valvular endocarditis (2,10,24,31,37,40). While several lines of evidence indicate a role for Fn in endovascular bacterial adherence during endocardial infections (21,30,42), other results strongly point to multifactorial events of bacterial adherence as the cause of heart valve damage (17).B. henselae Pap31 was originally described as a bacterial surface protein that is associated with the bacteriophage-like particle from B. henselae (6). It is possible that this protein is involved in packaging or phage particle assembly, or, alternatively, it may be merely a surface protein that copurifies with the particles. Pap31 is homologous to the multigene heparinbinding protein (encoded by hbp) family in Bartonella quintana, Neisseria opacity proteins (Opa), Brucella OMP31 (a putative porin), and Agrobacterium tumefaciens OMP25 (an immunogenic surface protein) (34). HbpA, the most prominent member of the Hbp family of proteins, has been reported to play a role as a hemin receptor for B. quintana, and Opa proteins in Neisseria gonorrhoeae promote adherence (4,8,20,50). The aims of the present study were to express and purify B. henselae Pap31; to characterize its binding to Fn, Hep, and host cells; and to investigate the effect of anti-Pap31 on B. henselae adherence to host cells.
MATERIALS AND METHODSBacteria, growth conditions, and reagents. B. henselae strain Houston-1 (ATCC 49882) was grown on tryptic soy plates supplemented with 5% sheep blood in a humidified atmosphere at 37°C under 5% CO 2 . Escherichia coli BL21(DE3)pLysS was grown in Luria-Bertani broth. The intact human Fn and Fn fragments (Fig. 1)