The role of HiTI, a serine protease inhibitor from Haematobia irritans irritans (Diptera : Muscidae) in the control of fly and bacterial proteases

The role of HiTI, a serine protease inhibitor from Haematobia irritans irritans (Diptera : Muscidae) in the control of fly and bacterial proteases

Author Azzolini, S. S. Google Scholar
Sasaki, S. D. Google Scholar
Campos, ITN Google Scholar
Torquato, RJS Google Scholar
Juliano, M. A. Google Scholar
Tanaka, A. S. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract Blood-sucking arthropods are vectors responsible for the transmission of several pathogens and parasites to vertebrate animals. the horn fly Haematobia irritans irritans (Diptera: Muscidae) and the tick Boophilus microplus are important hematophagous ecto-parasites that cause losses in cattle production. A serine protease inhibitor from a thorax extract of the fly H. irritans irritans (HiTI) was previously isolated, characterized and cloned. in the present study we described the expression, purification, and characterization of the recombinant HiTI (rHiTI) and its possible role in the control of different endogenous and bacterial proteases. rHiTI was successfully expressed using the pPIC9 expression vector with a yield of 4.2 mg/L of active rHiTI. the recombinant HiTI purified by affinity chromatography on trypsin-Sepharose had a molecular mass of 6.53 kDa as determined by LS-ESI mass spectrometry and inhibition constants (Kis) similar to those of native HiTI for bovine trypsin and human neutrophil elastase of 0.4 and 1.0 nM, respectively. Purified rHiTI also showed inhibitory activity against the trypsin-like enzyme of H. i. irritans using its possible natural substrates, fibrinogen and hemoglobin; and also inhibited the OmpT endoprotease of Escherichia coli using fluorogenic substrates. the present results confirm that HiTI may play a role in the control of fly endogenous proteases but also suggest a role in the inhibition of pathogen proteases. (c) 2005 Elsevier Inc. All rights reserved.
Keywords fly
serine protease inhibitor
innate immunity
omp-t endoprotease
Language English
Date 2005-09-01
Published in Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 111, n. 1, p. 30-36, 2005.
ISSN 0014-4894 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 30-36
Origin http://dx.doi.org/10.1016/j.exppara.2005.03.013
Access rights Closed access
Type Article
Web of Science ID WOS:000231354500005
URI http://repositorio.unifesp.br/handle/11600/28446

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