title: Allosteric Regulation of Fibronectin/α5β1 Interaction by Fibronectin-Binding MSCRAMMs.
abstract: Adherence of microbes to host tissues is a hallmark of infectious disease and is often mediated by a class of adhesins termed MSCRAMMs (Microbial Surface Components Recognizing Adhesive Matrix Molecules). Numerous pathogens express MSCRAMMs that specifically bind the heterodimeric human glycoprotein fibronectin (Fn). In addition to roles in adhesion, Fn-binding MSCRAMMs exploit physiological Fn functions. For example, several pathogens can invade host cells by a mechanism whereby MSCRAMM-bound Fn bridges interaction with α5β1 integrin. Here, we investigate two Fn-binding MSCRAMMs, FnBPA (Staphylococcus aureus) and BBK32 (Borrelia burgdorferi) to probe structure-activity relationships of MSCRAMM-induced Fn/α5β1integrin activation. Circular dichroism, fluorescence resonance energy transfer, and dynamic light scattering techniques uncover a conformational rearrangement of Fn involving domains distant from the MSCRAMM binding site. Surface plasmon resonance experiments demonstrate a significant enhancement of Fn/α5β1 integrin affinity in the presence of FnBPA or BBK32. Detailed kinetic analysis of these interactions reveal that this change in affinity can be attributed solely to an increase in the initial Fn/α5β1 on-rate and that this rate-enhancement is dependent on high-affinity Fn-binding by MSCRAMMs. These data implicate MSCRAMM-induced perturbation of specific intramolecular contacts within the Fn heterodimer resulting in activation by exposing previously cryptic α5β1 interaction motifs. By correlating structural changes in Fn to a direct measurement of increased Fn/α5β1 affinity, this work significantly advances our understanding of the structural basis for the modulation of integrin function by Fn-binding MSCRAMMs.
authors.affiliation: Center for Infectious and Inflammatory Diseases, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, 77030, United States of America.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Liang
authors.affiliation: Center for Infectious and Inflammatory Diseases, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, 77030, United States of America.
authors.forename: Brandon L
authors.initials: BL
authors.lastname: Garcia
authors.affiliation: Dep. of Molecular Medicine, UdR INSTM, Center for Tissue Engineering (C.I.T.), University of Pavia, 27100, Pavia, Italy.|Dep. of Occupational Medicine, Ergonomy and Disability, Salvatore Maugeri Foundation, IRCCS, Nanotechnology Laboratory, 27100, Pavia, Italy.
authors.forename: Livia
authors.initials: L
authors.lastname: Visai
authors.affiliation: Center for Infectious and Inflammatory Diseases, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, 77030, United States of America.
authors.forename: Sabitha
authors.initials: S
authors.lastname: Prabhakaran
authors.affiliation: Department of Biology, University of York, York, YO10 5DD, United Kingdom.
authors.forename: Nicola A G
authors.initials: NA
authors.lastname: Meenan
authors.affiliation: Department of Biology, University of York, York, YO10 5DD, United Kingdom.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Potts
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, M13 9PT, United Kingdom.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Humphries
authors.affiliation: Center for Infectious and Inflammatory Diseases, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, 77030, United States of America.
authors.forename: Magnus
authors.initials: M
authors.lastname: Höök
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics*; D000494:Allosteric Regulation / Q000235:genetics; D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D001665:Binding Sites; D025065:Borrelia burgdorferi / Q000235:genetics* / Q000472:pathogenicity; D002352:Carrier Proteins / Q000235:genetics*; D002942:Circular Dichroism; D005353:Fibronectins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D039081:Integrin alpha5beta1 / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D011485:Protein Binding; D013211:Staphylococcus aureus / Q000235:genetics* / Q000472:pathogenicity; D013329:Structure-Activity Relationship
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005353:Fibronectins; D039081:Integrin alpha5beta1; C509820:MSCRAMM proteins, Staphylococcus