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Title: Characterization of proteinases from the midgut of Rhipicephalus (Boophilus) microplus involved in the generation of antimicrobial peptides
Authors: Cruz, Carlos E.
Fogaca, Andrea C.
Nakayasu, Ernesto S.
Angeli, Claudia B.
Belmonte, Rodrigo
Almeida, Igor C.
Miranda, Antonio [UNIFESP]
Miranda, Maria Teresa M.
Tanaka, Aparecida Sadae [UNIFESP]
Braz, Gloria R.
Craik, Charles S.
Schneider, Eric
Caffrey, Conor R.
Daffre, Sirlei
Universidade de São Paulo (USP)
Univ Texas El Paso
Universidade Federal de São Paulo (UNIFESP)
Universidade Federal do Rio de Janeiro (UFRJ)
Univ Calif San Francisco
Issue Date: 27-Jul-2010
Publisher: Biomed Central Ltd
Citation: Parasites & Vectors. London: Biomed Central Ltd, v. 3, 15 p., 2010.
Abstract: Background: Hemoglobin is a rich source of biologically active peptides, some of which are potent antimicrobials (hemocidins). A few hemocidins have been purified from the midgut contents of ticks. Nonetheless, how antimicrobials are generated in the tick midgut and their role in immunity is still poorly understood. Here we report, for the first time, the contribution of two midgut proteinases to the generation of hemocidins.Results: An aspartic proteinase, designated BmAP, was isolated from the midgut of Rhipicephalus (Boophilus) microplus using three chromatographic steps. Reverse transcription-quantitative polymerase chain reaction revealed that BmAP is restricted to the midgut. the other enzyme is a previously characterized midgut cathepsin L-like cysteine proteinase designated BmCL1. Substrate specificities of native BmAP and recombinant BmCL1 were mapped using a synthetic combinatorial peptide library and bovine hemoglobin. BmCL1 preferred substrates containing non-polar residues at P2 subsite and polar residues at P1, whereas BmAP hydrolysed substrates containing non-polar amino acids at P1 and P1'.Conclusions: BmAP and BmCL1 generate hemocidins from hemoglobin alpha and beta chains in vitro. We postulate that hemocidins may be important for the control of tick pathogens and midgut flora.
ISSN: 1756-3305
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