A NOVEL RGD-BINDING MEMBRANE PROTEIN ON BLOOD PLATELETS

血小板上新型 RGD 结合膜蛋白

• 批准号: 2220156
• 负责人: Stephen CT Lam
• 金额: $ 11.64万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-05-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2220156
• 关键词: affinity chromatography; binding proteins; bioassay; blocking antibody; cell adhesion; cell adhesion molecules; chemical binding; crosslink; dimer; glycoproteins; human tissue; immunochemistry; laboratory rabbit; ligands; liposomes; membrane proteins; peptides; platelet aggregation; platelets; protein purification; protein sequence; protein structure function; radiotracer; receptor; receptor binding; synthetic peptide

Platelet adhesion to subendothelial constituents and platelet aggregation are important in hemostasis and thrombosis, and are mediated in part by the binding of 3 adhesive proteins which contain Arg-Gly-Asp (RGD) sequences: fibrinogen, fibronectin and von Willebrand factor. GPIIb-IIIa is a receptor for these adhesive proteins and binds to peptides containing the RGD sequence. Moreover, GPIIb-IIIa is a member of widely distributed adhesion receptor superfamily termed Integrins whose members are comprised of two non-identical alpha and beta subunits. Besides GPIIb-IIIa, certain other integrins also recognize the RGD sequence. We recently identified a platelet membrane protein of Mr approximately 160 kDa (P160) which binds to peptides containing the RGD sequence. This proposal will test the hypothesis that P160 forms a heterodimer complex with another platelet membrane protein to constitute a novel platelet integrin. To determine if P160 forms a heterodimer with a beta subunit, we will analyze the polypeptide content of immunoprecipitates formed with antibodies reactive with purified P160. In addition, we will examine the hydrodynamic properties of purified P160 to establish its Mr under non- denaturing conditions. Furthermore, we will investigate whether the sequence of P160 is similar to known integrins by direct sequencing of the amino-terminus and fragments of P160. Similar studies will be performed with the putative beta subunit of P160 should one be identified. The RGD-binding domain of P160 will be localized and characterized by affinity chromatography of proteolytic digests of purified P160, and by chemical crosslinking of RGD peptides to P160 followed by proteolytic cleavage. Since several membrane proteins which bind RGD serve adhesive functions, we will test the hypothesis that P160 plays a role in platelet adhesive reactions. Firstly, this will be accomplished by reconstituting purified P160 into radioactive liposomes and examining its binding to various RGD-containing adhesive proteins in a solid phase assay. Secondly, we will identify antibodies and RGD peptides which preferentially block the binding of P160 to immobilized RGD peptides. The capacity of these antibodies and peptides to inhibit platelet adhesive functions will be assessed. These studies will provide fundamental information about a membrane protein likely to be involved in mechanisms underlying platelet adhesion or aggregation.

血小板粘附到内皮下成分和血小板

项目成果

Immunolocalization of beta 1 integrins in platelets and platelet-derived microvesicles.

血小板和血小板衍生微泡中 β1 整合素的免疫定位。

• 发表时间: 1993
• 期刊: Blood
• 影响因子: 20.3
• 作者: Wencel-Drake,JD;Dieter,MG;Lam,SC
• 通讯作者: Lam,SC

Isolation and characterization of a chymotryptic fragment of platelet glycoprotein IIb-IIIa retaining Arg-Gly-Asp binding activity.

保留精氨酸-甘氨酸-天冬氨酸结合活性的血小板糖蛋白 IIb-IIIa 胰凝乳蛋白酶片段的分离和表征。

• 发表时间: 1992
• 期刊: The Journal of biological chemistry
• 作者: Lam,SC

The fibrinogen gamma chain dodecapeptide inhibits agonist-induced aggregation of rabbit platelets and fibrinogen binding to rabbit glycoprotein IIb-IIIa.

纤维蛋白原 γ 链十二肽抑制激动剂诱导的兔血小板聚集以及纤维蛋白原与兔糖蛋白 IIb-IIIa 的结合。

• 发表时间: 1999
• 期刊: Thrombosis and haemostasis.
• 作者: Rand,ML;Packham,MA;Taylor,DM;Yeo,EL;Gemmell,CH;Patil,S;Lam,SC
• 通讯作者: Lam,SC

Critical roles for the COOH-terminal NITY and RGT sequences of the integrin beta3 cytoplasmic domain in inside-out and outside-in signaling.

• DOI: 10.1083/jcb.200303120
• 发表时间: 2003-07-21
• 期刊: The Journal of cell biology
• 作者: Xi X;Bodnar RJ;Li Z;Lam SC;Du X
• 通讯作者: Du X

TA205, an anti-talin monoclonal antibody, inhibits integrin-talin interaction.

TA205 是一种抗 talin 单克隆抗体，可抑制整合素 - talin 相互作用。

• DOI: 10.1016/j.febslet.2006.02.077
• 发表时间: 2006
• 期刊: FEBS letters.
• 作者: Xing,Baodong;Thuppal,Shalini;Jedsadayanmata,Arom;Du,Xiaoping;Lam,StephenC-T
• 通讯作者: Lam,StephenC-T