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

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 colt and its activity was characterized. Ayadualin inhibited both collagen and ADP-induced platelet aggregations by interfering with the binding of integrin alpha(IIb)beta(3) 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. (C) 2015 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved

Molecular Diversity between Salivary Proteins from New World and Old World Sand Flies with Emphasis on Bichromomyia olmeca, the Sand Fly Vector of Leishmania mexicana in Mesoamerica

“…Recently, Kato and colleagues (2015) described a dual role for a Lu . ayacuchensis RGD containing protein called Ayadualin [ 32 ]. This protein inhibits platelet aggregation by binding to the integrin α IIb β 3 , an RGD-dependent function, and prevents blood coagulation by targeting FXII activation, an RGD-independent role [ 32 ].…”

rLj-RGD3 Suppresses the Growth of HeyA8 Cells in Nude Mice

“…At present, native RGD motif-containing peptides or proteins were mostly reported in the venom of snakes or the salivary glands of ticks, flies, tabanids, mosquitoes, and leeches, and not only suppressed the tumor growth and metastasis, but also inhibit platelet aggregation [ 20 , 21 , 22 , 23 , 24 , 25 ]. In 2010, a novel protein which contains three RGD motifs (Lj-RGD3) was first found in the buccal glands of L. japonica , one of the most primitive vertebrates [ 8 ].…”

Sergentomyia schwetzi: Salivary gland transcriptome, proteome and enzymatic activities in two lineages adapted to different blood sources

“…Sergentomyia schwetzi saliva RGD-containing peptides with an anti-platelet aggregation function [64] and several protein families with an unknown biological function, like the C-type lectins, 71 kDa salivary protein, spider-toxin-like and ML domain proteins [59,62,[65][66][67]. Even though the S. schwetzi is a sand fly species from OW, we identified in its salivary gland transcriptome salivary 5'-nucleotidase, 71 kDa salivary protein and the protein containing C-type lectin domain.…”