Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense
Trypanosoma brucei contain a serine oligopeptidase (OP-Tb) that is released into (and remains active in) the blood of trypanosome-infected animals. Here a similar enzyme from Trypanosoma congolense is described. This oligopeptidase, called OP-Tc, was purified using three-phase partitioning, and ion-...
| Autores principales: | , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
1999
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/29269 |
| _version_ | 1855526860467732480 |
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| author | Morty, R.E. Authié, Edith Troeberg, L. Lonsdale-Eccles, John D. Coetzer, T.H.T. |
| author_browse | Authié, Edith Coetzer, T.H.T. Lonsdale-Eccles, John D. Morty, R.E. Troeberg, L. |
| author_facet | Morty, R.E. Authié, Edith Troeberg, L. Lonsdale-Eccles, John D. Coetzer, T.H.T. |
| author_sort | Morty, R.E. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Trypanosoma brucei contain a serine oligopeptidase (OP-Tb) that is released into (and remains active in) the blood of trypanosome-infected animals. Here a similar enzyme from Trypanosoma congolense is described. This oligopeptidase, called OP-Tc, was purified using three-phase partitioning, and ion-exchange and affinity chromatography. OP-Tc is inhibited by alkylating agents, by serine peptidase-specific inhibitors including 3,4-dichloroisocoumarin, 4-(2-aminoethyl) benzenesulfonylfluoride and diispropylfluoro-phosphate and by other peptidase inhibitors including leupeptin, antipain and peptidyl chloromethyl ketones. Reducing agents such as dithiothreitol enhanced activity as did heparin, spermine and spermidine. The enzyme has trypsin-like specificity since it cleaved fluorogenic peptides that have basic amino acid residues (Arg or Lys) in the P1 position. Potential substrates without a basic residue in P1 were not hydrolysed. Although OP-Tc has weak arginine aminopeptidase activity, the enzyme clearly preferred substrates that had amino acids in the P2 and P3 positions. Overall. OP-Tc appears to be less efficient than OP-Tb because it usually displayed lower Kcat/Km values for the substrates tested. However, like OP-Tb, the best substrate for OP-Tc was Cbz-Arg-Arg-AMC (Km=0.72 M, Kcat=96 s-1). OP-Tc preference for amino acids in the P2 position was (Gly, Lys, Arg) > Phe >Leu >Pro. The results also suggest that the P3-binding site has hydrophobic characteristics. OP-Tc may not be a naturally immunodominant molecule because neither IgG nor IgM anti - OP-Tc antibodies were detected in the blood of experimentally infected cattle. |
| format | Journal Article |
| id | CGSpace29269 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 1999 |
| publishDateRange | 1999 |
| publishDateSort | 1999 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | CGSpace292692024-05-01T08:17:38Z Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense Morty, R.E. Authié, Edith Troeberg, L. Lonsdale-Eccles, John D. Coetzer, T.H.T. trypanosoma congolense peptides disease resistance enzymes antibodies trypsin Trypanosoma brucei contain a serine oligopeptidase (OP-Tb) that is released into (and remains active in) the blood of trypanosome-infected animals. Here a similar enzyme from Trypanosoma congolense is described. This oligopeptidase, called OP-Tc, was purified using three-phase partitioning, and ion-exchange and affinity chromatography. OP-Tc is inhibited by alkylating agents, by serine peptidase-specific inhibitors including 3,4-dichloroisocoumarin, 4-(2-aminoethyl) benzenesulfonylfluoride and diispropylfluoro-phosphate and by other peptidase inhibitors including leupeptin, antipain and peptidyl chloromethyl ketones. Reducing agents such as dithiothreitol enhanced activity as did heparin, spermine and spermidine. The enzyme has trypsin-like specificity since it cleaved fluorogenic peptides that have basic amino acid residues (Arg or Lys) in the P1 position. Potential substrates without a basic residue in P1 were not hydrolysed. Although OP-Tc has weak arginine aminopeptidase activity, the enzyme clearly preferred substrates that had amino acids in the P2 and P3 positions. Overall. OP-Tc appears to be less efficient than OP-Tb because it usually displayed lower Kcat/Km values for the substrates tested. However, like OP-Tb, the best substrate for OP-Tc was Cbz-Arg-Arg-AMC (Km=0.72 M, Kcat=96 s-1). OP-Tc preference for amino acids in the P2 position was (Gly, Lys, Arg) > Phe >Leu >Pro. The results also suggest that the P3-binding site has hydrophobic characteristics. OP-Tc may not be a naturally immunodominant molecule because neither IgG nor IgM anti - OP-Tc antibodies were detected in the blood of experimentally infected cattle. 1999-07 2013-06-11T09:22:59Z 2013-06-11T09:22:59Z Journal Article https://hdl.handle.net/10568/29269 en Limited Access Elsevier Molecular and Biochemical Parasitology;102: 144-155 |
| spellingShingle | trypanosoma congolense peptides disease resistance enzymes antibodies trypsin Morty, R.E. Authié, Edith Troeberg, L. Lonsdale-Eccles, John D. Coetzer, T.H.T. Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title | Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title_full | Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title_fullStr | Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title_full_unstemmed | Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title_short | Purification and characterisation of a typsin-like serine oligo peptidase from Trypanosoma congolense |
| title_sort | purification and characterisation of a typsin like serine oligo peptidase from trypanosoma congolense |
| topic | trypanosoma congolense peptides disease resistance enzymes antibodies trypsin |
| url | https://hdl.handle.net/10568/29269 |
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