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First published online January 3, 2006
Journal of Experimental Biology 209, 343-352 (2006)
Published by The Company of Biologists 2006
doi: 10.1242/jeb.02003

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Vasotab, a vasoactive peptide from horse fly Hybomitra bimaculata (Diptera, Tabanidae) salivary glands

Peter Taká^1,*,, Miles A. Nunn^2,*, János Mészáros³, Olga Pecháová⁴, Norbert Vrbjar⁵, Petra Vlasáková¹, Milan Kozánek¹, Mária Kazimírová¹, George Hart³, Patricia A. Nuttall² and Milan Labuda¹

¹ Institute of Zoology, Slovak Academy of Sciences, Dúbravská cesta 9, 845 06 Bratislava, Slovak Republic
² Centre of Ecology and Hydrology, NERC, Oxford, UK
³ Department of Medicine, University Clinical Departments, The Duncan Building, Daulby Street, Liverpool, L69 3GA, UK
⁴ Institute of Normal and Pathological Physiology
⁵ Institute for Heart Research, Department of Biochemistry, Slovak Academy of Sciences, Bratislava, Slovak Republic

Corresponding author (e-mail: uzaetaka{at}savba.sk)

Accepted 16 November 2005

Horse flies feed from superficial haematomas and probably rely heavily on the pharmacological properties of their saliva to find blood. Here we describe the first evidence of vasodilators in horse fly Hybomitra bimaculata (Diptera, Tabanidae) salivary gland extract and clone and express one of the active peptides (termed vasotab). Physiological tests using crude salivary gland extracts and reverse-phase HPLC fractions demonstrated positive inotropism in isolated rat hearts, vasodilatation of coronary and peripheral vessels, and Na, K-ATPase inhibition. One of the vasoactive fractions was analysed by N-terminal Edman degradation and a 47-amino-acid sequence obtained. A full-length cDNA encoding the peptide was cloned from a phage library using degenerate primer PCR and the peptide expressed in insect cells. A 20-amino-acid signal sequence precedes the mature 56-amino-acid vasotab peptide, which is a member of the Kazal-type protease inhibitor family. The peptide has a unique 7-amino-acid insertion between the third and fourth cysteine residues. The recombinant peptide prolonged the action potential and caused positive inotropism of isolated rat heart myocytes, and may be an ion channel modulator.

Key words: vasodilator, salivary gland, horse fly, Hybomitra bimaculata

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