title: Homozygous Cys542-->Arg substitution in GPIIIa in a Swiss patient with type I Glanzmann's thrombasthenia.
abstract: Glanzmann's thrombasthenia (GT) arises from a qualitative or quantitative defect in the GPIIb-IIIa complex (integrin alphaIIbbeta3), the mediator of platelet aggregation. We describe a patient in whom clinical and laboratory findings typical of type I GT were found together with a second pathology involving neurological and other complications symptomatic of tuberous sclerosis. Analysis of platelet proteins by Western blotting revealed trace amounts of normally migrating GPIIb and equally small amounts of GPIIIa of slightly slower than normal migration. Flow cytometry confirmed a much decreased binding to platelets of monoclonal antibodies to GPIIb, GPIIIa or GPIIb-IIIa, and an antibody to the alphav subunit also showed decreased binding. Nonradioactive PCR single-strand conformation polymorphism analysis followed by direct sequencing of PCR-amplified DNA fragments showed a homozygous point mutation (T to C) at nucleotide 1722 of GPIIIa cDNA and which led to a Cys542-->Arg substitution in the GPIIIa protein. The mutation gave rise to a HinP1 I restriction site in exon 11 of the GPIIIa gene and allele-specific restriction enzyme analysis of family members confirmed that a single mutated allele was inherited from each parent. This amino acid substitution presumably changes the capacity for disulphide bond formation within the cysteine-rich core region of GPIIIa and its study will provide new information on GPIIb-IIIa and alphavbeta3 structure and biosynthesis.
authors.affiliation: UMR 5533 CNRS, Hôpital Cardiologique, Pessac, France.
authors.forename: J
authors.initials: J
authors.lastname: Ruan
authors.forename: M
authors.initials: M
authors.lastname: Schmugge
authors.forename: K J
authors.initials: KJ
authors.lastname: Clemetson
authors.forename: E
authors.initials: E
authors.lastname: Cazes
authors.forename: R
authors.initials: R
authors.lastname: Combrie
authors.forename: F
authors.initials: F
authors.lastname: Bourre
authors.forename: A T
authors.initials: AT
authors.lastname: Nurden
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics*; D001120:Arginine / Q000235:genetics; D015153:Blotting, Western; D002675:Child, Preschool; D003545:Cysteine / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D006720:Homozygote; D006801:Humans; D010375:Pedigree; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013915:Thrombasthenia / Q000235:genetics*
chemical_list: D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D001120:Arginine; D003545:Cysteine