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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2007-2-27
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pubmed:abstractText |
Aspergillus fumigatus is an opportunistic fungal pathogen that infects immunocompromised patients. A putative aspartic protease gene (ctsD; 1425 bp; intron-free) was identified and cloned. CtsD is evolutionarily distinct from all previously identified A. fumigatus aspartic proteases. Recombinant CtsD was expressed in inclusion bodies in Escherichia coli (0.2mg/g cells) and subjected to extensive proteolysis in the baculovirus expression system. Activation studies performed on purified, refolded, recombinant CtsD resulted in protease activation with a pH(opt)4.0 and specific activity=10 U/mg. Pepstatin A also inhibited recombinant CtsD activity by up to 72% thereby confirming classification as an aspartic protease. Native CtsD was also immunologically identified in culture supernatants and purified from fungal cultures using pepstatin-agarose affinity chromatography (7.8 microg CtsD/g mycelia). In A. fumigatus, semi-quantitative RT-PCR analysis revealed expression of ctsD in minimal and proteinaceous media only. Expression of ctsD was absent under nutrient-rich conditions. Expression of ctsD was also detected, in vivo, in the Galleria mellonella virulence model following A. fumigatus infection.
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pubmed:grant |
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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 |
May
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pubmed:issn |
1046-5928
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
53
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
216-24
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:17275325-Amino Acid Motifs,
pubmed-meshheading:17275325-Amino Acid Sequence,
pubmed-meshheading:17275325-Animals,
pubmed-meshheading:17275325-Aspartic Acid Endopeptidases,
pubmed-meshheading:17275325-Aspergillus fumigatus,
pubmed-meshheading:17275325-Baculoviridae,
pubmed-meshheading:17275325-Base Sequence,
pubmed-meshheading:17275325-Binding Sites,
pubmed-meshheading:17275325-Chromatography, Affinity,
pubmed-meshheading:17275325-Cloning, Molecular,
pubmed-meshheading:17275325-Culture Media,
pubmed-meshheading:17275325-Enzyme Activation,
pubmed-meshheading:17275325-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:17275325-Escherichia coli,
pubmed-meshheading:17275325-Evolution, Molecular,
pubmed-meshheading:17275325-Gene Expression,
pubmed-meshheading:17275325-Genes, Fungal,
pubmed-meshheading:17275325-Hydrogen-Ion Concentration,
pubmed-meshheading:17275325-Hydrolysis,
pubmed-meshheading:17275325-Immunoglobulin G,
pubmed-meshheading:17275325-Inclusion Bodies,
pubmed-meshheading:17275325-Larva,
pubmed-meshheading:17275325-Models, Biological,
pubmed-meshheading:17275325-Molecular Sequence Data,
pubmed-meshheading:17275325-Moths,
pubmed-meshheading:17275325-Pepstatins,
pubmed-meshheading:17275325-Phylogeny,
pubmed-meshheading:17275325-Protein Folding,
pubmed-meshheading:17275325-Protein Sorting Signals,
pubmed-meshheading:17275325-Recombinant Proteins,
pubmed-meshheading:17275325-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:17275325-Survival Rate,
pubmed-meshheading:17275325-Virulence
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pubmed:year |
2007
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pubmed:articleTitle |
Isolation, activity and immunological characterisation of a secreted aspartic protease, CtsD, from Aspergillus fumigatus.
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pubmed:affiliation |
National Institute for Cellular Biotechnology, Department of Biology, National University of Ireland Maynooth, Co. Kildare, Ireland.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Research Support, N.I.H., Extramural
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