Bacterial adherence to the host tissue is recognized as a key step in the development of a bacterial infection. In the present proposal the extracellular connective tissue matrix rather than membrane components of the host cell is considered as a substrate for bacterial adherence and colonization. Recent studies in our and other laboratories which show that certain bacteria produce distinct cell wall components that specifically bind to fibronectin will be taken as a starting point for the proposed investigation. The structures of sites in the fibronectin molecule binding to model strains of Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli will be determined. To this end the protein will be cleaved by the use of proteolytic enzymes and CNBr. Generated peptides will subsequently be isolated and their ability to bind to bacteria or inhibit the binding of intact 125I-fibronectin to bacteria will be examined. The bacterial receptors responsible for fibronectin binding will be isolated and characterized. Different techniques will be tried to solubilize the fibronectin receptors which subsequently will be purified using affinity chromatography on Sepharose gels substituted with fibronectin or active fibronectin fragments in combination with conventional protein fractionation techniques. The isolated receptors will be characterized with respect to chemical and immunological properties. Specifically monoclonal antibodies will be raised against the receptor component and monoclonals that block the binding of I-fibronectin to bacteria will be searched for. The role of fibronectin in bacterial adherence will be examined. A model system consisting of coverslips coated with fibronectin or fibroblasts cultured in vitro will be used as substrates for adherence of fibronectin binding bacteria as well as receptor negative mutants. Attempts will be made to inhibit adherence by preincubating the bacteria with A) soluble fibronectin and its active fragments and B) monoclonal antibodies directed against the fibronectin receptors.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020624-04
Application #
3130402
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1984-12-01
Project End
1989-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Alabama Birmingham
Department
Type
School of Medicine & Dentistry
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Casillas-Ituarte, Nadia N; Cruz, Carlos H B; Lins, Roberto D et al. (2017) Amino acid polymorphisms in the fibronectin-binding repeats of fibronectin-binding protein A affect bond strength and fibronectin conformation. J Biol Chem 292:8797-8810
Garcia, Brandon L; Zhi, Hui; Wager, Beau et al. (2016) Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex. PLoS Pathog 12:e1005404
Arora, Srishtee; Uhlemann, Anne-Catrin; Lowy, Franklin D et al. (2016) A Novel MSCRAMM Subfamily in Coagulase Negative Staphylococcal Species. Front Microbiol 7:540
Ganesh, Vannakambadi K; Liang, Xiaowen; Geoghegan, Joan A et al. (2016) Lessons from the Crystal Structure of the S. aureus Surface Protein Clumping Factor A in Complex With Tefibazumab, an Inhibiting Monoclonal Antibody. EBioMedicine 13:328-338
Liang, Xiaowen; Garcia, Brandon L; Visai, Livia et al. (2016) Allosteric Regulation of Fibronectin/?5?1 Interaction by Fibronectin-Binding MSCRAMMs. PLoS One 11:e0159118
Kuipers, Annemarie; Stapels, Daphne A C; Weerwind, Lleroy T et al. (2016) The Staphylococcus aureus polysaccharide capsule and Efb-dependent fibrinogen shield act in concert to protect against phagocytosis. Microbiology 162:1185-94
Ko, Ya-Ping; Kang, Mingsong; Ganesh, Vannakambadi K et al. (2016) Coagulase and Efb of Staphylococcus aureus Have a Common Fibrinogen Binding Motif. MBio 7:e01885-15
Prasad, Joni M; Gorkun, Oleg V; Raghu, Harini et al. (2015) Mice expressing a mutant form of fibrinogen that cannot support fibrin formation exhibit compromised antimicrobial host defense. Blood 126:2047-58
Somarajan, Sudha R; La Rosa, Sabina Leanti; Singh, Kavindra V et al. (2015) The fibronectin-binding protein Fnm contributes to adherence to extracellular matrix components and virulence of Enterococcus faecium. Infect Immun 83:4653-61
Galloway-Peña, Jessica R; Liang, Xiaowen; Singh, Kavindra V et al. (2015) The identification and functional characterization of WxL proteins from Enterococcus faecium reveal surface proteins involved in extracellular matrix interactions. J Bacteriol 197:882-92

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