Inhibition of platelet aggregation of a mutant proinsulin chimera engineered by introduction of a native Lys-Gly-Asp-containing sequence.
An eight amino acid sequence, CAKGDWNC, from disintegrin barbourin, was introduced into an inactive human proinsulin molecule between the B28 and A2 sites to construct a chimeric, anti-thrombosis recombinant protein. The constructed Lys-Gly-Asp (KGD)-proinsulin gene was expressed in Escherichia coli and then purified. The KGD-proinsulin chimera protein inhibits human platelet aggregation, induced by ADP, with an IC50 value (molar concentration causing 50% inhibition of platelet aggregation) of 830 nM: and demonstrates also specific affinity to glycoprotein IIb/IIIa receptor. Its insulin receptor binding activity remains as low as 0.04% with native insulin as a control.
Jing
Jian
J
Department of Biochemistry and Biotechnology, Laboratory of Biotechnology and Protein Engineering, Beijing Normal University, 100875, Beijing, China. jjing@bnu.edu.cn
Lu
Shan
S
D000818:Animals; D002478:Cells, Cultured; D003435:Crotalid Venoms / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D017839:Crotalus; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000819:agonists; D011384:Proinsulin / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology
D003435:Crotalid Venoms; D009842:Oligopeptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011993:Recombinant Fusion Proteins; C068858:barbourin; D011384:Proinsulin