Purification, characterization, and cloning of a serine proteinase inhibitor from the ectoparasite Haematobia irritans irritans (Diptera: Muscidae)
| Autor: | Aparecida S. Tanaka, Simone S. Azzolini, Joyce M.C. Santos, Ricardo J.S. Torquato, Izaura Y. Hirata, Renato Andreotti, Adriana F. Souza |
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| Rok vydání: | 2003 |
| Předmět: |
DNA
Complementary Serine Proteinase Inhibitors Sequence analysis Immunology Molecular Sequence Data Biology Chromatography Affinity Serine Affinity chromatography Sequence Analysis Protein Complementary DNA Animals Amino Acid Sequence Cloning Molecular Peptide sequence Molecular mass Base Sequence Elastase Muscidae General Medicine biology.organism_classification Molecular biology Haematobia irritans Molecular Weight Infectious Diseases Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Chromatography Gel Parasitology Cattle |
| Zdroj: | Experimental parasitology. 106(3-4) |
| ISSN: | 0014-4894 |
| Popis: | The fly Haematobia irritans irritans 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 7029Da 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 0.20nM, 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. |
| Databáze: | OpenAIRE |
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