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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
34
|
| pubmed:dateCreated |
1994-1-4
|
| pubmed:abstractText |
We identified a new homozygous missense mutation His373-->Leu in the CYP17 gene of two sisters with 17 alpha-hydroxylase deficiency with an elevated plasma aldosterone concentration by sequencing their genomic DNAs amplified by polymerase chain reaction. Using polymerase chain reaction-based site-directed mutagenesis, we prepared a DNA that encoded the Leu373 mutant protein. COS-1 cells transfected with the mutant DNA, despite having an RNA hybridizable to the P450c17 cDNA, did not show 17 alpha-hydroxylase and 17,20-lyase activities. Also, the cells were devoid of 11 beta-hydroxylase and aldosterone synthase activities. To examine the mechanism by which the single amino acid change His373-->Leu eliminates activity, we expressed N-terminally modified P450c17 proteins with and without the Leu373 mutation in Escherichia coli and performed spectral studies. Membrane preparations from E. coli cells expressing the wild-type form of the modified enzyme showed an absorption peak at 449 nm upon addition of carbon monoxide in the reduced state and produced characteristic substrate-induced difference spectra, whereas those from the cells expressing the mutant form did not show these spectral changes. The 17 alpha-hydroxylase and 17,20-lyase activities were observed only in E. coli cells expressing the wild-type enzyme. These results show that the His373-->Leu mutant does not incorporate the heme prosthetic group properly and suggest a critical role of His373 in heme binding.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Codon,
http://linkedlifedata.com/resource/pubmed/chemical/DNA,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Primers,
http://linkedlifedata.com/resource/pubmed/chemical/Histidine,
http://linkedlifedata.com/resource/pubmed/chemical/Leucine,
http://linkedlifedata.com/resource/pubmed/chemical/Steroid 17-alpha-Hydroxylase
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Dec
|
| pubmed:issn |
0021-9258
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
5
|
| pubmed:volume |
268
|
| pubmed:geneSymbol |
CYP17
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
25811-7
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:8245018-Adolescent,
pubmed-meshheading:8245018-Adrenal Hyperplasia, Congenital,
pubmed-meshheading:8245018-Adult,
pubmed-meshheading:8245018-Alleles,
pubmed-meshheading:8245018-Amino Acid Sequence,
pubmed-meshheading:8245018-Animals,
pubmed-meshheading:8245018-Base Sequence,
pubmed-meshheading:8245018-Blotting, Southern,
pubmed-meshheading:8245018-Cell Line,
pubmed-meshheading:8245018-Codon,
pubmed-meshheading:8245018-DNA,
pubmed-meshheading:8245018-DNA Primers,
pubmed-meshheading:8245018-Exons,
pubmed-meshheading:8245018-Female,
pubmed-meshheading:8245018-Histidine,
pubmed-meshheading:8245018-Humans,
pubmed-meshheading:8245018-Leucine,
pubmed-meshheading:8245018-Leukocytes,
pubmed-meshheading:8245018-Male,
pubmed-meshheading:8245018-Molecular Sequence Data,
pubmed-meshheading:8245018-Mutagenesis, Site-Directed,
pubmed-meshheading:8245018-Point Mutation,
pubmed-meshheading:8245018-Polymerase Chain Reaction,
pubmed-meshheading:8245018-Restriction Mapping,
pubmed-meshheading:8245018-Steroid 17-alpha-Hydroxylase,
pubmed-meshheading:8245018-Transfection
|
| pubmed:year |
1993
|
| pubmed:articleTitle |
Mutation of histidine 373 to leucine in cytochrome P450c17 causes 17 alpha-hydroxylase deficiency.
|
| pubmed:affiliation |
Department of Internal Medicine, Toyama Medical and Pharmaceutical University Faculty of Medicine, Japan.
|
| pubmed:publicationType |
Journal Article,
Case Reports,
Research Support, Non-U.S. Gov't
|