Purification, characterization, and cloning of a serine proteinase inhibitor from the ectoparasite Haematobia irritans irritans (Diptera : Muscidae)

Purification, characterization, and cloning of a serine proteinase inhibitor from the ectoparasite Haematobia irritans irritans (Diptera : Muscidae)

Author Azzolini, S. S. Google Scholar
Santos, JMC Google Scholar
Souza, A. F. Google Scholar
Torquato, RJS Google Scholar
Hirata, I. Y. Google Scholar
Andreotti, R. Google Scholar
Tanaka, A. S. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
Abstract The fly Haematobia irritans irritant is one of the most important ectoparasites in cattle production, due to, its ability to suck large amounts of blood. This report describes the purification and characterization of a serine proteinase inhibitor (HiTI) present in H. i. irritans head and thorax extracts. the HiTI purified by, affinity chromatography on trypsin-Sepharose has a molecular mass of 7029 Da by MALDI-TOF mass spectrometry. HiTI inhibited bovine trypsin, human neutrophil elastase, and a trypsin-like enzyme, purified from H. i. irritans abdomen with dissociation constants of 0.57, 1.30, and 0.20 nM, respectively. the HiTI partial amino acid sequence allowed its classification into the BPTI-Kunitz-type family. An HiTI cDNA fragment was cloned in the pGEMT vector using RT-PCR. the translated amino acid sequence of HiTI cDNA confirmed a unique Kunitz-type-domain protein. Our results suggest that HiTI could control some endogenous enzyme, e.g., the H. i. irritans trypsin-like protein. (C) 2004 Elsevier Inc. All rights reserved.
Keywords Haematobia irritans irritans
Serine proteinase inhibitor
Kunitz-type inhibitor
HiTI
H. i. irritans trypsin inhibitor
bovine trypsin
human plasmin
human neutrophil elastase
bovine thrombin
human factor Xa
human factor XIIa
human plasma kallikrein
subtilisin A
MALDI-TOF
matrix assisted laser desorption ionization time-of-flight
cDNA
complementary deoxyribonucleic acid
RTPCR
reverse transcription polymerase chain reaction
TFA
trifluoroacetic acid
Language English
Date 2004-03-01
Published in Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 106, n. 3-4, p. 103-109, 2004.
ISSN 0014-4894 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 103-109
Origin http://dx.doi.org/10.1016/j.exppara.2004.01.012
Access rights Closed access
Type Article
Web of Science ID WOS:000221992300005
URI http://repositorio.unifesp.br/handle/11600/27646

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