Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense

The catalytic domains of two closely related cysteine proteinases (CP l and CP2) from Trypanosoma congolense, referred to as Cl and C2, were expressed as proforms in Escherichia coli (C 1) and in the baculovirus system (C I and C2). While the bacterial expression system did not allow recovery of act...

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Autores principales: Boulange, A.F., Serveau, C., Brillard, M., Minet, C., Gauthier, F., Diallo, A., Lalmanach, G., Authié, Edith
Formato: Journal Article
Lenguaje:Inglés
Publicado: Elsevier 2001
Materias:
Acceso en línea:https://hdl.handle.net/10568/32953
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author Boulange, A.F.
Serveau, C.
Brillard, M.
Minet, C.
Gauthier, F.
Diallo, A.
Lalmanach, G.
Authié, Edith
author_browse Authié, Edith
Boulange, A.F.
Brillard, M.
Diallo, A.
Gauthier, F.
Lalmanach, G.
Minet, C.
Serveau, C.
author_facet Boulange, A.F.
Serveau, C.
Brillard, M.
Minet, C.
Gauthier, F.
Diallo, A.
Lalmanach, G.
Authié, Edith
author_sort Boulange, A.F.
collection Repository of Agricultural Research Outputs (CGSpace)
description The catalytic domains of two closely related cysteine proteinases (CP l and CP2) from Trypanosoma congolense, referred to as Cl and C2, were expressed as proforms in Escherichia coli (C 1) and in the baculovirus system (C I and C2). While the bacterial expression system did not allow recovery of active C 1, the baculovirus system led to secretion of inactive zymogens which could be processed at acidic pH into mature enzymes. Active Cl and C2 were purified from serum-free culture supernatants by anion-exchange chromatography and characterised. Their kinetic parameters and pH activity profiles confirmed the relatedness between C2 and native CP2 (congopain). These properties also underline major functional differences between Cl and C2, that appear to relate to discrete but essential sequence differences. It is likely that these two enzymes perform distinct roles in vivo, in the parasite and/or in the host-parasite relationships.
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spelling CGSpace329532024-04-25T06:00:23Z Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense Boulange, A.F. Serveau, C. Brillard, M. Minet, C. Gauthier, F. Diallo, A. Lalmanach, G. Authié, Edith trypanosoma congolense cysteine peptidases baculoviridae infectious diseases parasitology The catalytic domains of two closely related cysteine proteinases (CP l and CP2) from Trypanosoma congolense, referred to as Cl and C2, were expressed as proforms in Escherichia coli (C 1) and in the baculovirus system (C I and C2). While the bacterial expression system did not allow recovery of active C 1, the baculovirus system led to secretion of inactive zymogens which could be processed at acidic pH into mature enzymes. Active Cl and C2 were purified from serum-free culture supernatants by anion-exchange chromatography and characterised. Their kinetic parameters and pH activity profiles confirmed the relatedness between C2 and native CP2 (congopain). These properties also underline major functional differences between Cl and C2, that appear to relate to discrete but essential sequence differences. It is likely that these two enzymes perform distinct roles in vivo, in the parasite and/or in the host-parasite relationships. 2001-11 2013-07-03T05:25:49Z 2013-07-03T05:25:49Z Journal Article https://hdl.handle.net/10568/32953 en Limited Access Elsevier International Journal for Parasitology;31(13): 1435-1440
spellingShingle trypanosoma congolense
cysteine
peptidases
baculoviridae
infectious diseases
parasitology
Boulange, A.F.
Serveau, C.
Brillard, M.
Minet, C.
Gauthier, F.
Diallo, A.
Lalmanach, G.
Authié, Edith
Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title_full Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title_fullStr Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title_full_unstemmed Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title_short Functional expression of the catalytic domains of two cysteine proteinases from Trypanosoma congolense
title_sort functional expression of the catalytic domains of two cysteine proteinases from trypanosoma congolense
topic trypanosoma congolense
cysteine
peptidases
baculoviridae
infectious diseases
parasitology
url https://hdl.handle.net/10568/32953
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