title: Natural and artificial mutations in αIIb integrin lead to a structural deformation of a calcium-binding site.
abstract: The platelet integrin αIIbβ3 is widely accepted as a structural and a functional model of the broad integrin protein family. The four calcium-binding sites in the αIIb subunit contribute to biogenesis and stability of the protein. Mansour et al. (J Thromb Haemost 9:192-200, 2011) showed that the natural Asn2Asp mutation causing Glanzmann thrombasthenia, prevented surface expression of αIIbβ3, whereas the artificial Asn2Gln mutation only decreased its level. Molecular dynamics simulations and EDTA chelation assay were used here to explore the mechanism of these structural deformations. We show a considerable expansion of the calcium-binding site 3 in Asn2Asp mutation, whereas the Asn2Gln toggles between normal and expanded conformations. The αIIbβ3 surface expression level correlates to the relative spending time in the expanded conformation. By a comparison to other calcium-binding sites of αIIb and of other α integrins we show that the size of a calcium-binding loop is conserved. EDTA chelation assay shows a sensitivity to calcium removal, which correlates with the reduction in αIIbβ3 surface expression and with the calcium binding site expansion, thus verifying the simulation data. Here we indicate that Asn2 mutation affects a calcium-binding site 3 of αIIb, which structural deformation is proposed to deprive calcium binding and interfere with an integrin intracellular trafficking and its surface expression.
authors.affiliation: The Amalia Biron Research Institute of Thrombosis and Haemostasis, Chaim Sheba Medical Center, 52621, Tel-Hashomer, Israel.
authors.forename: Wissam
authors.initials: W
authors.lastname: Mansour
authors.forename: Hagit
authors.initials: H
authors.lastname: Hauschner
authors.forename: Uri
authors.initials: U
authors.lastname: Seligsohn
authors.forename: Nurit
authors.initials: N
authors.lastname: Rosenberg
authors.forename: Yulia
authors.initials: Y
authors.lastname: Einav
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics*; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D006224:Cricetinae; D004492:Edetic Acid; D006860:Hydrogen Bonding; D056004:Molecular Dynamics Simulation; D009154:Mutation / Q000235:genetics / Q000502:physiology; D040201:Platelet Membrane Glycoprotein IIb / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D040201:Platelet Membrane Glycoprotein IIb; D004492:Edetic Acid; D002118:Calcium