Recombinant and native TviCATL from Trypanosoma vivax: Enzymatic characterisation and evaluation as a diagnostic target for animal African trypanosomosis.

Autor: Eyssen LE; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa., Vather P; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa., Jackson L; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa., Ximba P; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa., Biteau N; Laboratoire de Microbiologie Fondamentale et Pathogénicité, Université Bordeaux. UMR-CNRS 5234, 146, Rue Léo Saignat, 33076, Bordeaux Cedex, France., Baltz T; Laboratoire de Microbiologie Fondamentale et Pathogénicité, Université Bordeaux. UMR-CNRS 5234, 146, Rue Léo Saignat, 33076, Bordeaux Cedex, France., Boulangé A; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa; CIRAD, UMR INTERTRYP, 01009 Maputo, Mozambique; INTERTRYP, Univ Montpellier, CIRAD, IRD, Montpellier, France; Centro de Biotecnologia, Universidade Eduardo Mondlane, 01009 Maputo, Mozambique., Büscher P; Department of Biomedical Sciences, Institute of Tropical Medicine, Nationalestraat 155, B-2000, Antwerp, Belgium., Coetzer THT; Biochemistry, School of Life Sciences, University of KwaZulu-Natal (Pietermaritzburg campus), Private Bag X01, Scottsville, 3209, South Africa. Electronic address: coetzer@ukzn.ac.za.
Jazyk: angličtina
Zdroj: Molecular and biochemical parasitology [Mol Biochem Parasitol] 2018 Jul; Vol. 223, pp. 50-54. Date of Electronic Publication: 2018 Jul 07.
DOI: 10.1016/j.molbiopara.2018.07.001
Abstrakt: African animal trypanosomosis (nagana) is caused by tsetse-transmitted protozoan parasites. Their cysteine proteases are potential chemotherapeutic and diagnostic targets. The N-glycosylated catalytic domain of Trypanosoma vivax cathepsin L-like cysteine protease, rTviCATL cat , was recombinantly expressed and purified from culture supernatants while native TviCATL was purified from T. vivax Y486 parasite lysates. Typical of Clan CA, family C 1 proteases, TviCATL activity is sensitive to E-64 and cystatin and substrate specificity is defined by the S 2 pocket. Leucine was preferred in P 2 and basic and non-bulky, hydrophobic residues accepted in P 1 and P 3 respectively. Reversible aldehyde inhibitors, antipain, chymostatin and leupeptin, with Arg in P 1 and irreversible peptidyl chloromethylketone inhibitors with hydrophobic residues in P 2 inhibited TviCATL activity. TviCATL digested host proteins: bovine haemoglobin, serum albumin, fibrinogen and denatured collagen (gelatine) over a wide pH range, including neutral to slightly acidic pH. The recombinant catalytic domain of TviCATL showed promise as a diagnostic target for detecting T. vivax infection in cattle in an indirect antibody detection ELISA.
(Copyright © 2018 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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