title: Truncation of the cytoplasmic domain of beta3 in a variant form of Glanzmann thrombasthenia abrogates signaling through the integrin alpha(IIb)beta3 complex.
abstract: Glanzmann thrombasthenia is an inherited bleeding disorder characterized by absence or dysfunction of the platelet integrin alpha(IIb)beta3. Patient RM is a thrombasthenic variant whose platelets fail to aggregate in response to physiological agonists, despite the fact that they express abundant levels of alpha(IIb)beta3 on their surface. Binding of soluble fibrinogen or fibrinogen mimetic antibodies to RM platelets did not occur, except in the presence of ligand-induced binding site (LIBS) antibodies that transformed the RM integrin complex into an active conformation from outside the cell. Sequence analysis of PCR-amplified genomic DNA and platelet mRNA revealed a C2268T nucleotide substitution in the gene encoding the integrin beta3 subunit that resulted in an Arg724Ter mutation, producing a truncated protein containing only the first eight of the 47 amino acids normally present in the cytoplasmic domain. Functional analysis of both RM platelets and CHO cells stably expressing this truncated integrin revealed that the alpha(IIb)beta3Arg724Ter complex is able to mediate binding to immobilized fibrinogen, though downstream events, including cytoskeletally-mediated cell spreading and tyrosine phosphorylation of focal adhesion kinase, pp125FAK, fail to occur. These studies establish the importance of the membrane-distal portion of the integrin beta3 cytoplasmic domain in bidirectional transmembrane signaling in human platelets, and the role of integrin signaling in maintaining normal hemostasis in vivo.
authors.affiliation: Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee 53201-2178, USA.
authors.forename: R
authors.initials: R
authors.lastname: Wang
authors.forename: S J
authors.initials: SJ
authors.lastname: Shattil
authors.forename: D R
authors.initials: DR
authors.lastname: Ambruso
authors.forename: P J
authors.initials: PJ
authors.lastname: Newman
mesh_terms: D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D002462:Cell Membrane; D002648:Child; D003593:Cytoplasm / Q000648:ultrastructure; D006801:Humans; D039421:Integrin alpha2; D039661:Integrin beta3; D008024:Ligands; D008297:Male; D008969:Molecular Sequence Data; D015539:Platelet Activation; D010973:Platelet Adhesiveness; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000502:physiology*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins; D015398:Signal Transduction; D013329:Structure-Activity Relationship; D013915:Thrombasthenia / Q000235:genetics*
chemical_list: D015703:Antigens, CD; D039421:Integrin alpha2; D039661:Integrin beta3; D008024:Ligands; D010980:Platelet Membrane Glycoproteins; D011994:Recombinant Proteins