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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
28
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pubmed:dateCreated |
1991-11-8
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pubmed:databankReference |
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55915,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72400,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72401,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72402,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72403,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72404,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72405,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72406,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72407,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M72408,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/S57853
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pubmed:abstractText |
The complete amino acid sequence (296 amino acids) of Chromobacterium violaceum phenylalanine hydroxylase (PAH) was determined by nucleotide analysis of a DNA clone isolated using both a synthetic oligonucleotide probe based on the NH2-terminal amino acid sequence and an antibody against this enzyme. The ApaL I fragment (approximately 1.9 kilobase pairs) containing the entire PAH gene was subcloned in pBluescript II and induced by isopropyl-beta-D-thiogalactopyranoside. In order to eliminate fusion proteins the XbaI/ClaI fragment which contained the PAH gene from the Bluescript construct was subcloned into pMAC 5-8 containing the TAC promoter. The recombinant protein reacts with antibody raised to authentic C. violaceum PAH and its NH2-terminal 20-amino acid sequence and COOH-terminal amino acid residue were identical with the wild-type protein. Key physical and chemical characteristics of the recombinant protein, i.e. its copper content and Michaelis-Menten parameters, were the same as wild-type. Comparison of amino acid sequences revealed a highly conserved region between C. violaceum PAH and three different mammalian aromatic amino acid hydroxylases. This conserved area may well be a catalytically important domain of these pterin- and metal-requiring aromatic amino acid hydroxylases. The over-expression of C. violaceum PAH in Escherichia coli will facilitate the analysis of the enzyme mechanism by various spectroscopic methods.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
5
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pubmed:volume |
266
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
18454-9
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:1655752-Amino Acid Sequence,
pubmed-meshheading:1655752-Animals,
pubmed-meshheading:1655752-Base Sequence,
pubmed-meshheading:1655752-Chromobacterium,
pubmed-meshheading:1655752-Cloning, Molecular,
pubmed-meshheading:1655752-DNA, Bacterial,
pubmed-meshheading:1655752-Electron Spin Resonance Spectroscopy,
pubmed-meshheading:1655752-Escherichia coli,
pubmed-meshheading:1655752-Humans,
pubmed-meshheading:1655752-Molecular Sequence Data,
pubmed-meshheading:1655752-Phenylalanine Hydroxylase,
pubmed-meshheading:1655752-Rats,
pubmed-meshheading:1655752-Restriction Mapping,
pubmed-meshheading:1655752-Sequence Alignment
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pubmed:year |
1991
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pubmed:articleTitle |
Cloning and expression of Chromobacterium violaceum phenylalanine hydroxylase in Escherichia coli and comparison of amino acid sequence with mammalian aromatic amino acid hydroxylases.
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pubmed:affiliation |
Department of Chemistry, Pennsylvania State University, University Park 16802.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, Non-P.H.S.
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