title: Evidence that monoclonal antibodies directed against the integrin beta subunit plexin/semaphorin/integrin domain stimulate function by inducing receptor extension.
abstract: The overall structure of integrins is that of a ligand-binding head connected to two long legs. The legs can exhibit a pronounced bend at the "knees," and it has been proposed that the legs undergo a dramatic straightening when integrins transit from a low affinity to a high affinity state. The knee region contains domains from both alpha and beta subunits, including the N-terminal plexin/semaphorin/integrin (PSI) domain of the beta subunit. The role played by the knee domains in the regulation of integrin-ligand binding is uncertain. Here we show that: (i) monoclonal antibodies (mAbs) N29 and 8E3 have epitopes in the beta(1) subunit PSI domain and stimulate ligand binding to alpha(5)beta(1); (ii) N29 and 8E3 cause long range conformational changes that alter the ligand binding activity of the head region; (iii) the stimulatory action of these mAbs is dependent on the calf-1 domain, which forms part of the alpha subunit knee; and (iv) the epitopes of 8E3 and N29 map close to the extreme N terminus of the PSI and are likely to lie on the side of this domain that faces the alpha subunit. Taken together, our data suggest that the binding of these mAbs results in a levering apart of the PSI and calf-1 domains, and thereby causes the alpha and beta subunit knees to separate. Several major inferences can be drawn from our findings. First, the PSI domain appears to form part of an interface with the alpha subunit that normally restrains the integrin in a bent state. Second, the PSI domain is important for the transduction of conformational changes from the knee to head. Third, unbending is likely to provide a general mechanism for control of integrin-ligand recognition.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M13 9PT, United Kingdom. paul.mould@manchester.ac.uk
authors.forename: A Paul
authors.initials: AP
authors.lastname: Mould
authors.forename: Mark A
authors.initials: MA
authors.lastname: Travis
authors.forename: Stephanie J
authors.initials: SJ
authors.lastname: Barton
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Hamilton
authors.forename: Janet A
authors.initials: JA
authors.lastname: Askari
authors.forename: Susan E
authors.initials: SE
authors.lastname: Craig
authors.forename: Philip R
authors.initials: PR
authors.lastname: Macdonald
authors.forename: Richard A
authors.initials: RA
authors.lastname: Kammerer
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Buckley
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Humphries
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D016466:CHO Cells; D015815:Cell Adhesion Molecules / Q000737:chemistry*; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry; D005822:Genetic Vectors; D006801:Humans; D039081:Integrin alpha5beta1 / Q000378:metabolism; D039641:Integrin beta Chains / Q000737:chemistry*; D016023:Integrins / Q000737:chemistry; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D010920:Placenta / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D039961:Semaphorins / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D000939:Epitopes; D039081:Integrin alpha5beta1; D039641:Integrin beta Chains; D016023:Integrins; D008024:Ligands; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D039961:Semaphorins; C091529:plexin