A Ser162→ Leu Mutation within Glycoprotein (GP) IIIa (Integrin β3) Results in an Unstable αIIbβ3 Complex that Retains Partial Function in a Novel Form of Type II Glanzmann Thrombasthenia

Denise E. Jackson; Melanie M. White; Lisa K. Jennings; Peter J. Newman

doi:10.1055/s-0037-1615136

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1998; 80(01): 42-48
DOI: 10.1055/s-0037-1615136

Rapid Communication

Schattauer GmbH

A Ser₁₆₂→ Leu Mutation within Glycoprotein (GP) IIIa (Integrin β₃) Results in an Unstable α_IIbβ₃ Complex that Retains Partial Function in a Novel Form of Type II Glanzmann Thrombasthenia

Denise E. Jackson

¹Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI

,

Melanie M. White

²Departments of Medicine and Biochemistry, The University of Tennessee Memphis, Memphis, TN

,

Lisa K. Jennings

²Departments of Medicine and Biochemistry, The University of Tennessee Memphis, Memphis, TN

,

Peter J. Newman

¹Blood Research Institute, The Blood Center of Southeastern Wisconsin, Milwaukee, WI

³Departments of Cellular Biology and Pharmacology, Medical College of Wisconsin, Milwaukee, WI, USA

› Author Affiliations

Abstract

Summary

Platelets from Glanzmann thrombasthenia patient BL express approximately 30% of the normal α_IIbβ₃ content and support fibrin-mediated clot retraction, but fail to bind fibrinogen or aggregate following cellular activation. BL platelets bind neither activation-dependent nor activation-independent ligands. DNA sequence analysis of BL platelet mRNA revealed a homozygous C₅₈₃→T point mutation in a conserved region of β₃, resulting in a Ser162Leu amino acid substitution. This mutation appears to produce destabilizing effects on the α_IIbβ₃ complex, as evidenced by the fact that (1) the BL α_IIbβ₃ complex exhibited altered sedimentation velocity through sucrose gradients, (2) α_IIb and β₃ was not recognized by complex-dependent monoclonal antibodies or co-precipitated by integrin subunit-specific antibodies, and (3) biosynthesis and trafficking of the α_IIbβ₃Leu₁₆₂ complex was delayed relative to that of the wild-type control. Taken together, these data implicate the region encompassing Ser₁₆₂ in the stabilization and ligand binding properties of the α_IIbβ₃ complex.

Full Text

References

References
1 D’Souza SE, Haas TA, Piotrowicz R S, Byers-Ward V, McGrath DE, Soule HR, Cierniewski C, Plow EF, Smith JW. Ligand and cation binding are dual functions of a discrete segment of the integrin β₃ subunit: Cation displacement is involved in ligand binding.. Cell 1994; 79: 659-67.
2 D’Souza SE, Ginsberg MH, Matsueda GR, Plow EF. A discrete sequence in a platelet integrin is involved in ligand recognition.. Nature 1991; 350: 66-8.
3 Taylor DB, Gartner TK. A peptide corresponding to GPIIb 300-312, a presumptive fibrinogen γ-chain binding site on the platelet integrin GPIIb/IIIa, inhibits the adhesion of platelets to at least four adhesive ligands.. J Biol Chem 1992; 267: 11729-33.
4 Charo IF, Nannizzi L, Phillips DR, Hsu MA, Scarborough RM. Inhibition of fibrinogen binding to GPIIb-IIIa by a GPIIIa peptide.. J Biol Chem 1991; 266: 1415-21.
5 Lasz EC, McLane MA, Trybulec M, Kowalska MA, Khan S, Budzynski AZ, Niewiarowski S. β₃ integrin derived peptide 217-230 inhibits fibrinogen binding and platelet aggregation: significance of RGD sequences and fibrinogen A-chain.. Biochem Biophys Res Commun 1993; 190: 118-24.
6 Steiner B, Trzeciak A, Pfenninger G, Kouns WC. Peptides derived from a sequence within β₃ integrin bind to platelet α_IIbβ₃ (GPIIb-IIIa) and inhibit ligand binding.. J Biol Chem 1993; 268: 6870-73.
7 McKay BS, Annis DA, Honda S, Christie D, Kunicki TJ. Molecular requirements for assembly and function of a minimized human integrin α_IIbβ₃.. J Biol Chem 1996; 271: 30544-7.
8 Jennings LK, Phillips DR. Purification of glycoproteins IIb and III from human platelet plasma membranes and characterization of a calcium-dependent glycoprotein IIb-III complex.. J Biol Chem 1982; 257: 10458-66.
9 Kunicki T J, Pidard D, Rosa J-P, Nurden AT. The formation of Ca⁺⁺-depen-dent complexes of platelet membrane glycoproteins IIb and IIIa in solution as determined by crossed immunoelectrophoresis.. Blood 1981; 58: 268-78.
10 Bennett JS, Hoxie JA, Leitman SF, Vilaire G, Cines DB. Inhibition of fibrinogen binding to stimulated human platelets by a mAb.. Proc Natl Acad Sci USA 1983; 80: 2417-21.
11 Bennett JS, Vilaire G, Cines DB. Identification of the fibrinogen receptor on human platelets by photoaffinity labeling.. J Biol Chem 1982; 257: 8049-54.
12 Bray PF, Rosa J-P, Lingappa VR, Kan YW, McEver RP, Shuman MA. Biogenesis of the platelet receptor for fibrinogen: evidence for separate precursors for glycoproteins IIb and IIIa.. Proc Natl Acad Sci USA 1986; 83: 1480-4.
13 Duperray A, Troesch A, Berthier R, Chagnon E, Frachet P, Uzan G, Marguerie G. Biosynthesis and assembly of platelet GPIIb-IIIa in human megakaryocytes: Evidence that assembly between pro-GPIIb and GPIIIa is a prerequisite for expression of the complex on the cell surface.. Blood 1989; 74: 1603-11.
14 Duperray A, Berthier R, Chagnon E, Ryckewaert JJ, Ginsberg MH, Plow EF, Marguerie G. Biosynthesis and processing of platelet GPIIb-IIIa in human megakaryocytes.. J Cell Biol 1987; 104: 1665-73.
15 O’Toole TE, Loftus JC, Plow EF, Glass AA, Harper JR, Ginsberg MH. Efficient surface expression of platelet GPIIb-IIIa requires both subunits.. Blood 1989; 74: 14-8.
16 Rosa J-P, McEver RP. Processing and assembly of the integrin, glyco-protein IIb-IIIa in HEL cells.. J Biol Chem 1989; 264: 12596-603.
17 Glanzmann E. Hereditäre hämorrhagische Thrombasthenie: ein Beitrag zur Pathologie der Blutplättchen.. J Kinderkr 1918; 88: 113-41.
18 George JN, Caen JP, Nurden AT. Glanzmann’s thrombasthenia: the spectrum of clinical disease.. Blood 1990; 75: 1383-95.
19 Bennett JS, Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine.. J Clin Invest 1979; 64: 1393-401.
20 Caen JP. Glanzmann’s thrombasthenia.. Clin Haematol 1972; 1: 383-92.
21 Burk CD, Newman PJ, Lyman S, Gill J, Coller BS, Poncz M. A deletion in the gene for glycoprotein IIb associated with Glanzmann’s thrombasthenia.. J Clin Invest 1991; 87: 270-6.
22 Li L, Bray PF. Homologous recombination among three intragene Alu sequences causes an inversion-deletion resulting in the hereditary bleeding disorder Glanzmann thrombasthenia.. Am J Hum Genet 1995; 85: 140-9.
23 Peretz H, Rosenberg N, Usher S, Graff E, Newman PJ, Coller BS, Seligsohn U. Glanzmann’s thrombasthenia associated with deletion-insertion and alternative splicing in the glycoprotein IIb gene.. Blood 1994; 85: 414-20.
24 Iwamoto S, Nishiumi E, Kajii E, Ikemoto S. An exon 28 mutation resulting in alternative splicing of the glycoprotein IIb transcript and Glanzmann’s thrombasthenia.. Blood 1994; 83: 1017-23.
25 Newman PJ, Seligsohn U, Lyman S, Coller BS. The molecular genetic basis of Glanzmann thrombasthenia in the Iraqi-Jewish and Arab population in Israel.. Proc Natl Acad Sci USA 1991; 88: 3160-4.
26 Loftus JC, O’Toole TE, Plow EF, Glass A, Frelinger III AL, Ginsberg MH. A β₃ integrin mutation abolishes ligand binding and alters divalent cation-dependent conformation.. Science 1990; 249: 915-8.
27 Bajt ML, Ginsberg MH, Frelinger AL III, Berndt MC, Loftus JC. A spontaneous mutation of integrin α_IIbβ₃ (platelet glycoprotein IIb-IIIa) helps define a ligand binding site.. J Biol Chem 1992; 267: 3789-94.
28 Lanza F, Stierle A, Fournier D, Morales M, Andre G, Nurden AT, Cazenave J-P. A new variant of Glanzmann’s thrombasthenia (Strasbourg I). Platelets with functionally defective glycoprotein IIb-IIIa complexes and a glycoprotein IIIa ²¹⁴Arg→²¹⁴Trp mutation.. J Clin Invest 1992; 89: 1995-2004.
29 Poncz M, Rifat S, Coller BS, Newman PJ, Shattil SJ, Parrella T, Fortina P, Bennett JS. Glanzmann thrombasthenia secondary to a Gly²⁷³ → Asp mutation adjacent to the first calcium-binding domain of platelet glyco-protein IIb.. J Clin Invest 1994; 93: 172-9.
30 Chen Y-P, Djaffar I, Pidard D, Steiner B, Cieutat A-M, Caen JP, Rosa J-P. Ser⁷⁵² → Pro mutation in the cytoplasmic domain of integrin β₃ subunit and defective activation of the platelet integrin α_IIbβ₃ (glycoprotein IIb-IIIa) in a variant of Glanzmann thrombasthenia.. Proc Natl Acad Sci USA 1992; 89: 10169-73.
31 Wilcox DA, Wautier JL, Pidard D, Newman PJ. A single amino acid substitution flanking the fourth calcium binding domain of α_IIb prevents maturation of the α_IIbβ₃ integrin complex.. J Biol Chem 1994; 269: 4450-7.
32 Wilcox DA, Paddock CM, Lyman S, Gill JC, Newman PJ. Glanzmann thrombasthenia resulting from a single amino acid substitution between the second and third calcium-binding domains of GPIIb. Role of the GPIIb amino terminus in integrin subunit association.. J Clin Invest 1995; 95: 1553-60.
33 Wang RG, Ambruso DR, Newman PJ. Truncation in the cytoplasmic domain of integrin beta-3 subunit confers an inactivated state to the alpha-IIb-beta-3 complex in a variant type of Glanzmann thrombasthenia.. Blood 1994; 84: 244a.
34 Lee J-O, Rieu P, Arnaout MA, Liddington R. Crystal structure of the A domain from the α subunit of integrin CR3 (CD11b/CD18).. Cell 1995; 80: 631-8.
35 Nurden AT, Rosa J-P, Fournier D, Legrand C, Didry C, Parquet A, Pidard A. A variant of Glanzmann’s thrombasthenia with abnormal glycoprotein IIb-IIIa complexes in the platelet membrane.. J Clin Invest 1987; 79: 962-9.
36 Fournier DJ, Kabral A, Castaldi PA, Berndt MC. A variant of Glanzmann’s thrombasthenia characterized by abnormal glycoprotein IIb/IIIa complex formation.. Thromb Haemost 1989; 62 (03) 977-83.
37 Newman PJ, Weyerbusch-Bottum S, Visentin GP, Gidwitz S, White II GC. Type II Glanzmann thrombasthenia due to a destabilizing amino acid substitution in platelet membrane glycoprotein IIIa.. Thromb Haemost 1993; 69 (06) 1691a.
38 Jennings LK, Wang WC, Jackson CW, Fox CF, Bell A. Hemostasis in Glanzmann’s thrombasthenia (GT): GT platelets interfere with the aggregation of normal platelets.. Amer J Pediatr Hematol 1991; 13 (01) 84-90.
39 Kouns WC, Wall CD, White MM, Fox CF, Jennings LK. A conformation-dependent epitope of human platelet glycoprotein IIIa.. J Biol Chem 1990; 265: 20594-601.
40 Jennings LK, White MM, Mandrell TD. Interspecies comparison of platelet aggregation, LIBS expression and clot retraction: observed differences in GPIIb-IIIa functional activity.. Thromb Haemost 1995; 74 (06) 1551-6.
41 McEver RP, Bennett EM, Martin MN. Identification of two structurally and functionally distinct sites on human platelet membrane glycoprotein IIb-IIIa using monoclonal antibodies.. J Biol Chem 1983; 258: 5269-75.
42 Coller BS, Peerschke EI, Seligsohn U, Scudder LE, Nurden AT, Rosa JP. Studies on the binding of an alloimmune and two murine monoclonal antibodies to the platelet glycoprotein IIb-IIIa complex receptor.. J Lab Clin Med 1986; 107: 384-93.
43 Coller BS. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of platelet GPIIb-IIIa complex.. J Clin Invest 1985; 76: 101-8.
44 Weisel JW, Nagaswami C, Vilaire G, Bennett JS. Examination of the platelet membrane glycoprotein IIb-IIIa complex and its interaction with fibrinogen and other ligands by electron microscopy.. J Biol Chem 1992; 267: 16637-43.
45 Pidard D, Montgomery RR, Bennett JS, Kunicki TJ. Interaction of AP-2, a monoclonal antibody specific for the human platelet glycoprotein IIb-IIIa complex, with intact platelets.. J Biol Chem 1983; 258: 12582-6.
46 Coller BS, Peerschke EI, Scudder LE, Sullivan CA. Studies with a murine monoclonal antibody that abolishes ristocetin-induced binding of von Willebrand factor to platelets: Additional evidence in support of GPIb as a platelet receptor for von Willebrand factor.. Blood 1983; 61: 99-100.
47 Newman PJ, Allen RW, Kahn RA, Kunicki TJ. Quantitation of membrane glycoprotein IIIa on intact human platelets using the monoclonal antibody, AP-3.. Blood 1985; 65: 227-32.
48 Newman PJ, Gorski J, White GC, Gidwitz S, Cretney CJ, Aster RH. Enzymatic amplification of platelet-specific messenger RNA using the polymerase chain reaction.. J Clin Invest 1988; 82: 739-43.
49 Jennings LK, Ashmun RA, Wang WC, Dockter ME. Analysis of human platelet glycoproteins IIb-IIIa and Glanzmann’s thrombasthenia in whole blood by flow cytometry.. Blood 1986; 68 (01) 173-9.
50 Lefebvre P, White MD, Krumweide I, Cohen I. Role of actin in platelet function.. Eur J Cell Biol 1993; 62: 194-204.
51 Schoenwaelder SM, Jackson SP, Yuan Y, Teasdale MS, Salem HH, Mitchell CA. Tyrosine kinases regulate the cytoskeletal attachment of integrin α_IIbβ₃ (platelet glycoprotein IIb/IIIa) and cellular retraction of fibrin polymers.. J Biol Chem 1994; 269: 32479-87.
52 Schielen WJG, Voskuilen M, Tesser GI, Nieuwenhuizen W. The sequence Aα-(148-160) in fibrin, but not in fibrinogen, is accessible to monoclonal antibodies.. Proc Natl Acad Sci USA 1989; 86: 8951-4.
53 Ugarova TP, Budzynski AZ, Shattil SJ, Ruggeri ZM, Ginsberg MH, Plow EF. Conformational changes in fibrinogen elicited by its interaction with the platelet membrane glycoprotein GPIIb-IIIa.. J Biol Chem 1993; 268: 21080-7.
54 Taylor DB, Derrick JM, Gartner TK. Antibodies to GPIIbα (300-312) inhibit fibrinogen binding, clot retraction, and platelet adhesion to multiple ligands.. Proc Soc Exp Biol Med 1994; 205: 35-43.
55 Farrell DH, Thiagarajan P, Chung DW, Davie EW. Role of fibrinogen and chain sites in platelet aggregation.. Proc Natl Acad Sci 1992; 89: 10729-32.
56 Holmback K, Danton MJS, Suh TT, Daugherty CC, Degen JL. Impaired platelet aggregation and sustained bleeding in mice lacking the fibrinogen motif bound by integrin α_IIbβ₃.. EMBO J 1996; 15: 5760-71.
57 Rooney MM, Parise LV, Lord ST. Dissecting clot retraction and platelet aggregation.. J Biol Chem 1996; 271: 8553-5.