Structure-Guided Design of a High-Affinity Platelet Integrin α
 IIb
 β
 3
 Receptor Antagonist That Disrupts Mg
 2+
 Binding to the MIDAS

Zhu, Jieqing; Choi, Won-Seok; McCoy, Joshua G.; Negri, Ana; Zhu, Jinghua; Naini, Sarasija; Li, Jihong; Shen, Min; Huang, Wenwei; Bougie, Daniel W.; Rasmussen, Mark A.; Aster, Richard H.; Thomas, Craig J.; Filizola, Marta; Springer, Timothy A.; Coller, Barry S.

doi:10.1126/scitranslmed.3003576

Cited by 85 publications

(94 citation statements)

References 55 publications

(124 reference statements)

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“…[33][34][35] Using PAR4-AP-induced IP 3 generation, an important PAR4 signaling molecule, 11,36 overexpression of the PAR4 variants demonstrated differing signaling capacities. A potential limitation of these experiments is that 293 cell type-specific effects may not apply to platelets.…”

Section: Discussionmentioning

confidence: 99%

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

Edelstein¹,

Simon

Lindsay³

et al. 2014

Blood

115

131

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• White individuals have a high frequency of the common PAR4 gene (F2RL3) variant Ala120; blacks have a high frequency of Thr120.• PAR4 Thr120 induces greater signaling and is associated with greater platelet aggregation and reduced inhibition by a PAR4 antagonist.Human platelets express 2 thrombin receptors: protease-activated receptor (PAR)-1 and PAR4. Recently, we reported 3.7-fold increased PAR4-mediated aggregation kinetics in platelets from black subjects compared with white subjects. We now show that platelets from blacks (n 5 70) express 14% more PAR4 protein than those from whites (n 5 84), but this difference is not associated with platelet PAR4 function. Quantitative trait locus analysis identified 3 common single nucleotide polymorphisms in the PAR4 gene (F2RL3) associated with PAR4-induced platelet aggregation. Among these single nucleotide polymorphisms, rs773902 determines whether residue 120 in transmembrane domain 2 is an alanine (Ala) or threonine (Thr). Compared with the Ala120 variant, Thr120 was more common in black subjects than in white subjects (63% vs 19%), was associated with higher PAR4-induced human platelet aggregation and Ca 21 flux, and generated greater inositol 1,4,5-triphosphate in transfected cells. A second, less frequent F2RL3 variant, Phe296Val, was only observed in blacks and abolished the enhanced PAR4-induced platelet aggregation and 1,4,5-triphosphate generation associated with PAR4-Thr120. PAR4 genotype did not affect vorapaxar inhibition of platelet PAR1 function, but a strong pharmacogenetic effect was observed with the PAR4-specific antagonist YD-3 [1-benzyl-3(ethoxycarbonylphenyl)-indazole]. These findings may have an important pharmacogenetic effect on the development of new PAR antagonists. (Blood. 2014;124(23):3450-3458)

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Section: Discussionmentioning

confidence: 99%

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

Edelstein¹,

Simon

Lindsay³

et al. 2014

Blood

115

131

View full text Add to dashboard Cite

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“…The large-scale integrin conformational rearrangements, including the changing from a bent to an extended, and from a closed to an open headpiece conformation, have been directly visualized by electron microscopy (EM) (Takagi et al, 2002;Nishida et al, 2006;Ye et al, 2010;Eng et al, 2011;Zhu et al, 2012), crystallography (Xiao et al, 2004;Zhu et al, 2013), and small angle X-ray scattering (Mould et al, 2003;Eng et al, 2011). Integrin conformational change was also demonstrated on the cell surface by the exposure of epitopes (known as ligand-induced-binding sites, LIBS) that are masked in the inactive state (Frelinger et al, 1991;Sims et al, 1991;Humphries, 2004;Byron et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

The dual structural roles of the membrane distal region of α integrin cytoplasmic tail in integrin inside-out activation

Liu¹,

Wang²,

Thinn³

et al. 2015

Journal of Cell Science

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BSTRACTStudies on the mechanism of integrin inside-out activation have been focused on the role of b-integrin cytoplasmic tails, which are relatively conserved and bear binding sites for the intracellular activators including talin and kindlin. Cytoplasmic tails for a-integrins share a conserved GFFKR motif at the membrane-proximal region and this forms a specific interface with the b-integrin membraneproximal region to keep the integrin inactive. The a-integrin membrane-distal regions, after the GFFKR motif, are diverse both in length and sequence and their roles in integrin activation have not been well-defined. In this study, we report that the a-integrin cytoplasmic membrane-distal region contributes to maintaining integrin in the resting state and to integrin inside-out activation.Complete deletion of the a-integrin membrane-distal region diminished talin-and kindlin-mediated integrin ligand binding and conformational change. A proper length and suitable amino acids in a-integrin membrane-distal region was found to be important for integrin inside-out activation. Our data establish an essential role for the a-integrin cytoplasmic membrane-distal region in integrin activation and provide new insights into how talin and kindlin induce the high-affinity integrin conformation that is required for fully functional integrins.

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“…The biochemical and molecular abnormalities in GT have been studied for nearly 40 y (4,6,(8)(9)(10). In the past 10 y, high-resolution crystallography, electron microscopy, and computational studies of the αIIbβ3 receptor have provided atomic-level information on the correlation between receptor structure and function (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21). In addition, ethnic groups with relatively high prevalence of GT have been defined that share the same genetic abnormality based on founder mutations, and the dates that some of the mutations entered the population have been estimated (22)(23)(24)(25)(26)(27)(28).…”

mentioning

confidence: 99%

αIIbβ3 variants defined by next-generation sequencing: Predicting variants likely to cause Glanzmann thrombasthenia

Buitrago

Rendon²,

Liang

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Next-generation sequencing is transforming our understanding of human genetic variation but assessing the functional impact of novel variants presents challenges. We analyzed missense variants in the integrin αIIbβ3 receptor subunit genes ITGA2B and ITGB3 identified by whole-exome or -genome sequencing in the ThromboGenomics project, comprising ∼32,000 alleles from 16,108 individuals. We analyzed the results in comparison with 111 missense variants in these genes previously reported as being associated with Glanzmann thrombasthenia (GT), 20 associated with alloimmune thrombocytopenia, and 5 associated with aniso/macrothrombocytopenia. We identified 114 novel missense variants in ITGA2B (affecting ∼11% of the amino acids) and 68 novel missense variants in ITGB3 (affecting ∼9% of the amino acids). Of the variants, 96% had minor allele frequencies (MAF) < 0.1%, indicating their rarity. Based on sequence conservation, MAF, and location on a complete model of αIIbβ3, we selected three novel variants that affect amino acids previously associated with GT for expression in HEK293 cells. αIIb P176H and β3 C547G severely reduced αIIbβ3 expression, whereas αIIb P943A partially reduced αIIbβ3 expression and had no effect on fibrinogen binding. We used receiver operating characteristic curves of combined annotationdependent depletion, Polyphen 2-HDIV, and sorting intolerant from tolerant to estimate the percentage of novel variants likely to be deleterious. At optimal cut-off values, which had 69-98% sensitivity in detecting GT mutations, between 27% and 71% of the novel αIIb or β3 missense variants were predicted to be deleterious. Our data have implications for understanding the evolutionary pressure on αIIbβ3 and highlight the challenges in predicting the clinical significance of novel missense variants.Glanzmann | integrin | single-nucleotide variants | next-generation sequencing | molecular modeling

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Structure-Guided Design of a High-Affinity Platelet Integrin α _IIb β ₃ Receptor Antagonist That Disrupts Mg ²⁺ Binding to the MIDAS

Abstract: A new drug blocks platelet aggregation by interacting with the integrin receptor differently from current drugs that cause adverse side effects.

Cited by 85 publications

References 55 publications

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

The dual structural roles of the membrane distal region of α integrin cytoplasmic tail in integrin inside-out activation

αIIbβ3 variants defined by next-generation sequencing: Predicting variants likely to cause Glanzmann thrombasthenia

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Structure-Guided Design of a High-Affinity Platelet Integrin α IIb β 3 Receptor Antagonist That Disrupts Mg 2+ Binding to the MIDAS

Abstract: A new drug blocks platelet aggregation by interacting with the integrin receptor differently from current drugs that cause adverse side effects.

Cited by 85 publications

References 55 publications

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

Common variants in the human platelet PAR4 thrombin receptor alter platelet function and differ by race

The dual structural roles of the membrane distal region of α integrin cytoplasmic tail in integrin inside-out activation

αIIbβ3 variants defined by next-generation sequencing: Predicting variants likely to cause Glanzmann thrombasthenia

Contact Info

Product

Resources

About

Structure-Guided Design of a High-Affinity Platelet Integrin α _IIb β ₃ Receptor Antagonist That Disrupts Mg ²⁺ Binding to the MIDAS