Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida)
This study investigated protease activities in a crude extract and in vitro excretion/secretion (E/S) products of Philasterides dicentrarchi , a ciliate fish parasite causing economically significant losses in aquaculture. Gelatin/SDS–PAGE analysis (pH 4, reducing conditions) detected 7 bands with g...
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Cambridge University Press (CUP)
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Online Access: | http://dx.doi.org/10.1017/s0031182004004883 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182004004883 |
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crcambridgeupr:10.1017/s0031182004004883 2024-09-15T18:40:01+00:00 Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) PARAMÁ, A. IGLESIAS, R. ÁLVAREZ, M. F. LEIRO, J. UBEIRA, F. M. SANMARTÍN, M. L. 2004 http://dx.doi.org/10.1017/s0031182004004883 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182004004883 en eng Cambridge University Press (CUP) https://www.cambridge.org/core/terms Parasitology volume 128, issue 5, page 541-548 ISSN 0031-1820 1469-8161 journal-article 2004 crcambridgeupr https://doi.org/10.1017/s0031182004004883 2024-07-17T04:04:25Z This study investigated protease activities in a crude extract and in vitro excretion/secretion (E/S) products of Philasterides dicentrarchi , a ciliate fish parasite causing economically significant losses in aquaculture. Gelatin/SDS–PAGE analysis (pH 4, reducing conditions) detected 7 bands with gelatinolytic activity (approximate molecular weights 30–63 kDa) in the crude extract. The banding pattern observed in analysis of E/S products was practically identical, except for 1 low-molecular-weight band detected in the crude extract but not in the E/S products. In assays with synthetic peptide p -nitroanilide substrates, the crude extract hydrolysed substrates characteristic of cysteine proteases, namely Z-Arg-Arg pNA, Bz-Phe-Val-Arg pNA and Z-Phe-Arg pNA. These activities were strongly inhibited by the cysteine protease inhibitor E-64 and by Ac-Leu-Val-Lys aldehyde, a potent inhibitor of cysteine proteases of the cathepsin B protease subfamily. The proteases present in the crude extract degraded both type-I collagen and haemoglobin in vitro , consistent with roles in tissue invasion and nutrition respectively. Again, E-64 completely (collagen) or markedly (haemoglobin) inhibited this degradation. Finally, the histolytic activity of the ciliate in turbot fibroblast monolayers was strongly reduced in the presence of E-64, confirming the importance of secreted cysteine proteinases in the biology of Philasterides dicentrarchi . Article in Journal/Newspaper Turbot Cambridge University Press Parasitology 128 5 541 548 |
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Open Polar |
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Cambridge University Press |
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language |
English |
description |
This study investigated protease activities in a crude extract and in vitro excretion/secretion (E/S) products of Philasterides dicentrarchi , a ciliate fish parasite causing economically significant losses in aquaculture. Gelatin/SDS–PAGE analysis (pH 4, reducing conditions) detected 7 bands with gelatinolytic activity (approximate molecular weights 30–63 kDa) in the crude extract. The banding pattern observed in analysis of E/S products was practically identical, except for 1 low-molecular-weight band detected in the crude extract but not in the E/S products. In assays with synthetic peptide p -nitroanilide substrates, the crude extract hydrolysed substrates characteristic of cysteine proteases, namely Z-Arg-Arg pNA, Bz-Phe-Val-Arg pNA and Z-Phe-Arg pNA. These activities were strongly inhibited by the cysteine protease inhibitor E-64 and by Ac-Leu-Val-Lys aldehyde, a potent inhibitor of cysteine proteases of the cathepsin B protease subfamily. The proteases present in the crude extract degraded both type-I collagen and haemoglobin in vitro , consistent with roles in tissue invasion and nutrition respectively. Again, E-64 completely (collagen) or markedly (haemoglobin) inhibited this degradation. Finally, the histolytic activity of the ciliate in turbot fibroblast monolayers was strongly reduced in the presence of E-64, confirming the importance of secreted cysteine proteinases in the biology of Philasterides dicentrarchi . |
format |
Article in Journal/Newspaper |
author |
PARAMÁ, A. IGLESIAS, R. ÁLVAREZ, M. F. LEIRO, J. UBEIRA, F. M. SANMARTÍN, M. L. |
spellingShingle |
PARAMÁ, A. IGLESIAS, R. ÁLVAREZ, M. F. LEIRO, J. UBEIRA, F. M. SANMARTÍN, M. L. Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
author_facet |
PARAMÁ, A. IGLESIAS, R. ÁLVAREZ, M. F. LEIRO, J. UBEIRA, F. M. SANMARTÍN, M. L. |
author_sort |
PARAMÁ, A. |
title |
Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
title_short |
Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
title_full |
Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
title_fullStr |
Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
title_full_unstemmed |
Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi(Ciliophora: Scuticociliatida) |
title_sort |
cysteine proteinase activities in the fish pathogen philasterides dicentrarchi(ciliophora: scuticociliatida) |
publisher |
Cambridge University Press (CUP) |
publishDate |
2004 |
url |
http://dx.doi.org/10.1017/s0031182004004883 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182004004883 |
genre |
Turbot |
genre_facet |
Turbot |
op_source |
Parasitology volume 128, issue 5, page 541-548 ISSN 0031-1820 1469-8161 |
op_rights |
https://www.cambridge.org/core/terms |
op_doi |
https://doi.org/10.1017/s0031182004004883 |
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Parasitology |
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128 |
container_issue |
5 |
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541 |
op_container_end_page |
548 |
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1810484336002072576 |