1. Characterization of a novel cystatin type 2 from Rhipicephalus microplus midgut.
- Author
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Cardoso THS, Lu S, Gonzalez BRG, Torquato RJS, and Tanaka AS
- Subjects
- Animals, Cattle, Gene Expression, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins isolation & purification, Arthropod Proteins biosynthesis, Arthropod Proteins chemistry, Arthropod Proteins genetics, Arthropod Proteins isolation & purification, Intestinal Mucosa metabolism, Rhipicephalus chemistry, Rhipicephalus genetics, Rhipicephalus metabolism, Salivary Cystatins biosynthesis, Salivary Cystatins chemistry, Salivary Cystatins genetics, Salivary Cystatins isolation & purification
- Abstract
The Rhipicephalus (Boophilus) microplus is an exclusive bovine ectoparasite responsible for the transmission of pathogens that decrease meat, leather and milk productions. Cattle vaccination is an alternative to control tick infestations, but the discovery of potential antigens is still a challenge for researchers. Recently, our group performed a midgut transcriptome of engorged R. microplus tick, and out of 800 ESTs sequences one cystatin-coding sequence was identified and named Rmcystatin-4. In order to understand the physiological role of Rmcystatin-4, the aim of this work was the expression, purification and functional characterization of a novel type 2 cystatin from the tick R. microplus. Rmcystatin-4 gene expression was identified mostly in tick midgut suggesting its possible role in blood digestion control. Our data showed that rRmcystatin-4 was successfully expressed in active form using Pichia pastoris system and the purified inhibitor presented high selectivity to BmCl-1 (Ki = 0.046 nM). Moreover, rRmcystatin-4 was able to impaired BmCl-1 activity towards bovine hemoglobin., (Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.)
- Published
- 2017
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