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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 |
2000-10-19
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pubmed:abstractText |
The design of chimeric proteins is a major field of interest in structural biology and biotechnology. The successful design of the chimeric protein composed by the minimized reactive site domain of the low-molecular-mass trypsin inhibitor from Brassica napus (var. oleifera) seed (Ser3-Lys35; mini-RTI-III) and murine dihydrofolate reductase (DHFR) is reported here. The DHFR-mini-RTI-III chimeric protein was expressed in Escherichia coli, purified by metal-chelate affinity chromatography and oxidatively refolded. The affinity of the purified and refolded DHFR-mini-RTI-III for bovine trypsin (K = 5.0 x 10(-10) M) was closely similar to that determined for native RTI-III (K = 2.9 x 10(-10) M), at pH 8.2 and 22.0 degrees C. DHFR-mini-RTI-III may be regarded as a tool in structure-function studies and for developing multifunctional and multidomain proteinase inhibitors.
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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 |
Sep
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pubmed:issn |
0006-291X
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pubmed:author |
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pubmed:copyrightInfo |
Copyright 2000 Academic Press.
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pubmed:issnType |
Print
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pubmed:day |
7
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pubmed:volume |
275
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
817-20
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pubmed:dateRevised |
2005-11-17
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pubmed:meshHeading |
pubmed-meshheading:10973804-Amino Acid Motifs,
pubmed-meshheading:10973804-Amino Acid Sequence,
pubmed-meshheading:10973804-Animals,
pubmed-meshheading:10973804-Binding Sites,
pubmed-meshheading:10973804-Brassica,
pubmed-meshheading:10973804-Cattle,
pubmed-meshheading:10973804-Chromatography, Affinity,
pubmed-meshheading:10973804-Escherichia coli,
pubmed-meshheading:10973804-Kinetics,
pubmed-meshheading:10973804-Mice,
pubmed-meshheading:10973804-Molecular Sequence Data,
pubmed-meshheading:10973804-Plant Proteins,
pubmed-meshheading:10973804-Protein Engineering,
pubmed-meshheading:10973804-Protein Folding,
pubmed-meshheading:10973804-Recombinant Fusion Proteins,
pubmed-meshheading:10973804-Tetrahydrofolate Dehydrogenase,
pubmed-meshheading:10973804-Trypsin,
pubmed-meshheading:10973804-Trypsin Inhibitors
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pubmed:year |
2000
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pubmed:articleTitle |
A chimeric mini-trypsin inhibitor derived from the oil rape proteinase inhibitor type III.
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pubmed:affiliation |
Dipartimento di Genetica e Biologia Molecolare 'Charles Darwin', Università di Roma 'La Sapienza', Piazzale A. Moro 5, Rome, I-00185, Italy.
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pubmed:publicationType |
Journal Article
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