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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
3
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pubmed:dateCreated |
2008-3-3
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pubmed:databankReference |
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pubmed:abstractText |
Kudoa paniformis and Kudoa thyrsites (Myxozoa: Myxosporea) infections are associated with severe proteolysis of host muscle tissue post-mortem. The present study was undertaken to identify and characterize the protease responsible for myoliquefaction and determine mechanisms controlling protease function in vivo. N-terminal sequence analysis of partially purified protease from hake muscle infected with K. paniformis and K. thyrsites revealed a 23 amino acid sequence that aligned with cysteine proteases. Enzyme inhibition assays confirmed the presence of an essential active site cysteine residue. Using the above K. paniformis amino acid sequence data, a corresponding cDNA sequence from K. thyrsites plasmodia was elucidated revealing a cathepsin L proenzyme (Kth-CL). The translated amino acid sequence lacked a signal sequence characteristic of lysosomal and secreted proteins suggesting a unique cytoplasmic location. Only the proenzyme form of Kth-CL was present in Atlantic salmon muscle anti-mortem but this form became processed in vivo when infected muscle was stored at 4 degrees C. The proenzyme of Kth-CL showed uninhibited activity at pH 6.0, negligible activity at pH 6.5 and no measurable activity at pH 7.0 whilst the processed protease showed stability and function over a broad pH range (pH 4.5-8.8). The pH dependent processing and function of Kth-CL was consistent with histidine residues in the proregion playing a critical role in the regulation of Kth-CL.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
1096-4959
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
149
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
477-89
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:18187354-Amino Acid Sequence,
pubmed-meshheading:18187354-Animals,
pubmed-meshheading:18187354-Cathepsin L,
pubmed-meshheading:18187354-Cathepsins,
pubmed-meshheading:18187354-Chromatography, High Pressure Liquid,
pubmed-meshheading:18187354-Cysteine Endopeptidases,
pubmed-meshheading:18187354-Cytoplasm,
pubmed-meshheading:18187354-DNA, Complementary,
pubmed-meshheading:18187354-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:18187354-Eukaryota,
pubmed-meshheading:18187354-Fluorescent Antibody Technique,
pubmed-meshheading:18187354-Gadiformes,
pubmed-meshheading:18187354-Hydrogen-Ion Concentration,
pubmed-meshheading:18187354-Molecular Sequence Data,
pubmed-meshheading:18187354-Muscle, Skeletal,
pubmed-meshheading:18187354-Muscle Proteins,
pubmed-meshheading:18187354-Protease Inhibitors,
pubmed-meshheading:18187354-Protozoan Infections, Animal,
pubmed-meshheading:18187354-Salmo salar,
pubmed-meshheading:18187354-Sequence Analysis, DNA
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pubmed:year |
2008
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pubmed:articleTitle |
Identification, characterization and deduced amino acid sequence of the dominant protease from Kudoa paniformis and K. thyrsites: a unique cytoplasmic cysteine protease.
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pubmed:affiliation |
BC Centre for Aquatic Health Sciences, 621 Island Hwy, PO Box 277, Campbell River, BC Canada V9W 5B1. val.funk@cahs-bc.ca
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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