title: Binding Affinity of Metal Ions to the CD11b A-domain Is Regulated by Integrin Activation and Ligands.
abstract: The divalent cations Mg(2+) and Ca(2+) regulate the interaction of integrins with their cognate ligands, with Mg(2+) uniformly facilitating and Ca(2+) generally inhibiting such interactions in vitro. Because both cations are present in mm concentrations in vivo, the physiologic relevance of the in vitro observations is unclear. We measured the affinity of both cations to the inactive and active states of the ligand- and cation-binding A-domain (CD11bA) from integrin CD11b/CD18 in the absence and presence of the single-chain 107 antibody (scFv107), an activation-insensitive ligand-mimetic antibody. Using titration calorimetry, we found that Mg(2+) and Ca(2+) display equivalent (mm) affinities to inactive CD11bA. Activation induced a approximately 10-fold increase in the binding affinity of Mg(2+) to CD11bA with no change in that of Ca(2+) (106 microm +/- 16 and 2.1 mm +/- 0.19, respectively, n = 4). This increase is largely driven by favorable enthalpy. scFv107 induced a 50-80-fold increase in the binding affinity of Ca(2+) (but not Mg(2+) or Mn(2+)) to either form of CD11bA. Thus the affinity of metal ions to integrins is itself regulated by the activation state of these receptors and by certain ligands. These findings, which we expect will be applicable in vivo, elucidate a new level of regulation of the integrin-metal-ligand ternary complex and help explain some of the discrepant effects of Ca(2+) on integrin-ligand interactions.
authors.affiliation: Leukocyte Biology and Inflammation Program, Renal Unit, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129, USA.
authors.forename: Kaouther
authors.initials: K
authors.lastname: Ajroud
authors.forename: Takashi
authors.initials: T
authors.lastname: Sugimori
authors.forename: Wolfgang H
authors.initials: WH
authors.lastname: Goldmann
authors.forename: Dahmani M
authors.initials: DM
authors.lastname: Fathallah
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Xiong
authors.forename: M Amin
authors.initials: MA
authors.lastname: Arnaout
mesh_terms: D039481:CD11b Antigen / Q000737:chemistry / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D008024:Ligands; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D039481:CD11b Antigen; D008024:Ligands; D008670:Metals; D011994:Recombinant Proteins; D008345:Manganese; D008274:Magnesium; D002118:Calcium