A minimized human integrin alpha(5)beta(1) that retains ligand recognition.
Two isolated recombinant fragments from human integrin alpha(5)beta(1) encompassing the FG-GAP repeats III to VII of alpha(5) and the insertion-type domain from beta(1), respectively, are structurally well defined in solution, based on CD evidence. Divalent cation binding induces a conformational adaptation that is achieved by Ca(2+) or Mg(2+) (or Mn(2+)) with alpha(5) and only by Mg(2+) (or Mn(2+)) with beta(1). Mn(2+) bound to beta(1) is highly hydrated ( approximately 3 water molecules), based on water NMR relaxation, in agreement with a metal ion-dependent adhesion site-type metal coordination. Each fragment saturated with Mg(2+) (or Mn(2+)) binds a recombinant fibronectin ligand in an RGD-dependent manner. A conformational rearrangement is induced on the fibronectin ligand upon binding to the alpha(5), but not to the beta(1) fragment, based on CD. Ligand binding results in metal ion displacement from beta(1). Both alpha(5) and beta(1) fragments form a stable heterodimer (alpha(5)beta(1) mini-integrin) that retains ligand recognition to form a 1:1:1 ternary complex, in the presence of Mg(2+), and induces a specific conformational adaptation of the fibronectin ligand. A two-site model for RGD binding to both alpha and beta integrin components is inferred from our data using low molecular weight RGD mimetics.
Banères
J L
JL
Chimie Biomoléculaire et Interactions Biologiques, UPRESA CNRS 5074, Faculté de Pharmacie, 15 Av. Ch. Flahault, 34060 Montpellier Cédex 2, France. baneres@pharma.univ-montp1.fr
Roquet
F
F
Martin
A
A
Parello
J
J
D002118:Calcium / Q000378:metabolism; D002412:Cations / Q000378:metabolism*; D002942:Circular Dichroism; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008274:Magnesium / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese / Q000378:metabolism; D013058:Mass Spectrometry; D008956:Models, Chemical; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017469:Receptors, Fibronectin / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D002412:Cations; D005353:Fibronectins; D008024:Ligands; D009842:Oligopeptides; D010446:Peptide Fragments; D017469:Receptors, Fibronectin; D011994:Recombinant Proteins; D008345:Manganese; C047981:arginyl-glycyl-aspartic acid; D008274:Magnesium; D002118:Calcium