title: Construction and characterization of novel hirulog variants with antithrombin and antiplatelet activities.
abstract: The RGD sequence was used to design potent hirudin isoform 3 mimetic peptides with both antithrombin activity and antiplatelet aggregation activity. The RGD and proline were inserted between the catalytic active binding domain (D-Phe-Pro-Arg-Pro) on the N-terminus and the anion-binding exosite binding domain (QGDFEPIPEDAYDE) on the Cterminus. Thrombin titration assay and ATP-induced platelet aggregation test revealed that the peptide with the linker RGDWP or RGDGP possessed potent antithrombin and antiplatelet activities, while other peptides without the Pro residue in the linker only showed antithrombin activity. Similar results were obtained in the RGD-containing hirulog-1 variants. Our study indicates that the inserted Pro residue facilitates the exposure of RGD and the binding of the peptide to glycoprotein IIb/IIIa (GPIIb/IIIa). The strategy of combining the RGD sequence and the Pro residue may be used for future designs of bifunctional antithrombotic agents.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Yu
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Huang
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Dai
authors.forename: Mingxin
authors.initials: M
authors.lastname: Dong
authors.forename: Zhuguo
authors.initials: Z
authors.lastname: Liu
authors.forename: Shuo
authors.initials: S
authors.lastname: Yu
authors.forename: Jie
authors.initials: J
authors.lastname: Hu
authors.affiliation: Beijing Institute of Biotechnology, Beijing 100071, China. qy_dai@yahoo.com.
authors.forename: Qiuyun
authors.initials: Q
authors.lastname: Dai
mesh_terms: D000818:Animals; D058833:Antithrombin Proteins / Q000138:chemical synthesis*; D000991:Antithrombins / Q000138:chemical synthesis* / Q000737:chemistry; D020134:Catalytic Domain; D006629:Hirudins / Q000138:chemical synthesis / Q000737:chemistry*; D039661:Integrin beta3 / Q000378:metabolism; D008297:Male; D009842:Oligopeptides; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry*; D010975:Platelet Aggregation Inhibitors / Q000138:chemical synthesis*; D040201:Platelet Membrane Glycoprotein IIb / Q000378:metabolism; D020033:Protein Isoforms / Q000138:chemical synthesis / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000737:chemistry; D014914:Whole Blood Coagulation Time
chemical_list: D058833:Antithrombin Proteins; D000991:Antithrombins; D006629:Hirudins; D039661:Integrin beta3; D009842:Oligopeptides; D010446:Peptide Fragments; D010975:Platelet Aggregation Inhibitors; D040201:Platelet Membrane Glycoprotein IIb; D020033:Protein Isoforms; D011994:Recombinant Proteins; C047981:arginyl-glycyl-aspartic acid; C074619:bivalirudin