title: A naturally occurring mutation near the amino terminus of alphaIIb defines a new region involved in ligand binding to alphaIIbbeta3.
abstract: Decreased expression of functional alphaIIbbeta3 complexes on the platelet surface produces Glanzmann thrombasthenia. We have identified mutations of alphaIIb(P145) in 3 ethnically distinct families affected by Glanzmann thrombasthenia. Affected Mennonite and Dutch patients were homozygous and doubly heterozygous, respectively, for a P(145)A substitution, whereas a Chinese patient was doubly heterozygous for a P(145)L substitution. The mutations affect expression levels of surface alphaIIbbeta3 receptors on their platelets, which was confirmed by co-transfection of alphaIIb(P145A) and beta3 cDNA constructs in COS-1 cells. Each mutation also impaired the ability of alphaIIbbeta3 on affected platelets to interact with ligands. Moreover, when alphaIIb(P145A) and beta3 were stably coexpressed in Chinese hamster ovary cells, alphaIIbbeta3 was readily detected on the cell surface, but the cells were unable to adhere to immobilized fibrinogen or to bind soluble fluorescein isothiocyanate-fibrinogen after alphaIIbbeta3 activation by the activating monoclonal antibody PT25-2. Nonetheless, incubating affected platelets with the peptide LSARLAF, which binds to alphaIIb, induced PF4 secretion, indicating that the mutant alphaIIbbeta3 retained the ability to mediate outside-in signaling. These studies indicate that mutations involving alphaIIb(P145 )impair surface expression of alphaIIbbeta3 and that the alphaIIb(P145A) mutation abrogates ligand binding to the activated integrin. A comparative analysis of other alphaIIb mutations with a similar phenotype suggests that these mutations may cluster into a single region on the surface of the alphaIIb and may define a domain influencing ligand binding. (Blood. 2000;95:180188)
authors.affiliation: Departments of Pediatrics and Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
authors.forename: R B
authors.initials: RB
authors.lastname: Basani
authors.forename: D L
authors.initials: DL
authors.lastname: French
authors.forename: G
authors.initials: G
authors.lastname: Vilaire
authors.forename: D L
authors.initials: DL
authors.lastname: Brown
authors.forename: F
authors.initials: F
authors.lastname: Chen
authors.forename: B S
authors.initials: BS
authors.lastname: Coller
authors.forename: J M
authors.initials: JM
authors.lastname: Derrick
authors.forename: T K
authors.initials: TK
authors.lastname: Gartner
authors.forename: J S
authors.initials: JS
authors.lastname: Bennett
authors.forename: M
authors.initials: M
authors.lastname: Poncz
mesh_terms: D000293:Adolescent; D000328:Adult; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D001792:Blood Platelets / Q000502:physiology; D016466:CHO Cells; D019556:COS Cells; D002675:Child, Preschool; D002681:China / Q000208:ethnology; D002835:Christianity; D006224:Cricetinae; D005006:Ethnicity / Q000235:genetics; D005260:Female; D006579:Heterozygote; D006801:Humans; D008024:Ligands; D046911:Macromolecular Substances; D008297:Male; D008958:Models, Molecular; D009426:Netherlands / Q000208:ethnology; D010414:Pennsylvania; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017354:Point Mutation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000378:metabolism; D013915:Thrombasthenia / Q000235:genetics*; D014162:Transfection; D044465:White People / Q000235:genetics
chemical_list: D008024:Ligands; D046911:Macromolecular Substances; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011994:Recombinant Proteins