Ayadualin, a novel RGD peptide with dual antihemostatic activities from the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis

Biochimie. 2015 May:112:49-56. doi: 10.1016/j.biochi.2015.02.011. Epub 2015 Feb 24.

Abstract

Sequence analysis of the Lutzomyia (Lu.) ayacuchensis salivary gland cDNA library identified a short peptide containing an RGD (Arg-Gly-Asp) sequence flanked by two cysteine residues in the C-terminal end as the most abundant transcript. In the present study, a recombinant protein of the RGD-containing peptide, designated ayadualin, was expressed in Escherichia coli and its activity was characterized. Ayadualin inhibited both collagen and ADP-induced platelet aggregations by interfering with the binding of integrin αIIbβ3 to fibrinogen. The RGD sequence and cysteine residues located on both sides of the RGD sequence were essential for the inhibitory action. Moreover, ayadualin efficiently inhibited the intrinsic blood coagulation pathway irrespective of the RGD sequence. Measuring the enzymatic activity of coagulation factors using chromogenic substrates revealed that ayadualin efficiently inhibited factor XIIa (FXIIa) activity in a dose-dependent manner. In addition, pre-incubation of ayadualin with FXII inhibited FXIIa activity, while activated FXIIa was not affected by ayadualin, indicating that ayadualin inhibits the activation of FXII, but not enzymatic activity of FXIIa. These results indicated that ayadualin plays an important role in the blood feeding of Lu. ayacuchensis by inhibiting host hemostasis via dual mechanisms.

Keywords: Anticoagulant; Antiplatelet; RDG peptide; Saliva; Sand fly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Coagulation / drug effects*
  • Dose-Response Relationship, Drug
  • Factor XIIa / antagonists & inhibitors*
  • Humans
  • Insect Proteins / chemistry
  • Insect Proteins / pharmacology*
  • Insect Vectors / chemistry*
  • Leishmaniasis, Cutaneous*
  • Oligopeptides / chemistry
  • Oligopeptides / pharmacology*
  • Psychodidae / chemistry*

Substances

  • Insect Proteins
  • Oligopeptides
  • arginyl-glycyl-aspartic acid
  • Factor XIIa