Inhibition of platelet aggregation of a mutant proinsulin chimera engineered by introduction of a native Lys-Gly-Asp-containing sequence.

Abstract

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.

• Authors: Jian Jing, Shan Lu
• Journal: Biotechnology letters (Biotechnol Lett) Vol. 27 Issue 17 Pg. 1259-65 (Sep 2005) ISSN: 0141-5492 [Print] Netherlands
• PMID: 16215822 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Crotalid Venoms
• Oligopeptides
• Platelet Glycoprotein GPIIb-IIIa Complex
• Recombinant Fusion Proteins
• barbourin
• Proinsulin

Topics

• Animals
• Cells, Cultured
• Crotalid Venoms (chemistry, genetics, pharmacology)
• Crotalus
• Dose-Response Relationship, Drug
• Humans
• Oligopeptides (chemistry, genetics, pharmacology)
• Platelet Aggregation (drug effects)
• Platelet Glycoprotein GPIIb-IIIa Complex (agonists)
• Proinsulin (chemistry, genetics, pharmacology)
• Protein Engineering
• Protein Structure, Tertiary
• Recombinant Fusion Proteins (chemistry, genetics, pharmacology)