Source:http://linkedlifedata.com/resource/pubmed/id/18211827
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
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| pubmed:dateCreated |
2008-2-15
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| pubmed:abstractText |
Recently, S-adenosylhomocysteine hydrolase deficiency was confirmed for the first time in an adult. Two missense mutations in codons 89 (A>V) and 143 (Y>C) in the AdoHcyase gene were identified [N.R.M. Buist, B. Glenn, O. Vugrek, C. Wagner, S. Stabler, R.H. Allen, I. Pogribny, A. Schulze, S.H. Zeisel, I. Bari?, S.H. Mudd, S-Adenosylhomocysteine hydrolase deficiency in a 26-year-old man, J. Inh. Metab. Dis. 29 (2006) 538-545]. Accordingly, we have proven the Y143C mutation to be highly inactivating [R. Beluzi?, M. Cuk, T. Pavkov, K. Fumi?, I. Bari?, S.H. Mudd, I. Jurak, O. Vugrek, A single mutation at tyrosine 143 of human S-adenosylhomocysteine hydrolase renders the enzyme thermosensitive and effects the oxidation state of bound co-factor NAD, Biochem. J. 400 (2006) 245-253]. Now we report that the A89V exchange leads to a 70% loss of enzymatic activity, respectively. Circular dichroism analysis of recombinant p.A89V protein shows a significantly reduced unfolding temperature by 5.5 degrees C compared to wild-type. Gel filtration of mutant protein is almost identical to wild-type indicating assembly of subunits into the tetrameric complex. However, electrophoretic mobility of p.A89V is notably faster as shown by native polyacrylamide gel electrophoresis implicating changes to the overall charge of the mutant complex. 'Bioinformatics' analysis indicates that Val(89) collides with Thr(84) causing sterical incompatibility. Performing site-directed mutagenesis changing Thr(84) to 'smaller' Ser(84) but preserving similar physico-chemical properties restores most of the catalytic capabilities of the mutant p.A89V enzyme. On the other hand, substitution of Thr(84) with Lys(84) or Gln(84), thereby introducing residues with higher volume in proximity to Ala(89) results in inactivation of wild-type protein. In view of our mutational analysis, we consider changes in charge and the sterical incompatibility in mutant p.A89V protein as main reason for enzyme malfunction with AdoHcyase deficiency as consequence.
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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 |
Mar
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| pubmed:issn |
1090-2104
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| pubmed:author | |
| pubmed:issnType |
Electronic
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| pubmed:day |
28
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| pubmed:volume |
368
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
30-6
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| pubmed:meshHeading |
pubmed-meshheading:18211827-Adenosylhomocysteinase,
pubmed-meshheading:18211827-Catalytic Domain,
pubmed-meshheading:18211827-Chromatography, Gel,
pubmed-meshheading:18211827-Codon,
pubmed-meshheading:18211827-Humans,
pubmed-meshheading:18211827-Kinetics,
pubmed-meshheading:18211827-Models, Molecular,
pubmed-meshheading:18211827-Mutagenesis, Site-Directed,
pubmed-meshheading:18211827-Protein Structure, Tertiary,
pubmed-meshheading:18211827-Recombinant Proteins
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| pubmed:year |
2008
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| pubmed:articleTitle |
S-Adenosylhomocysteine hydrolase (AdoHcyase) deficiency: enzymatic capabilities of human AdoHcyase are highly effected by changes to codon 89 and its surrounding residues.
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| pubmed:affiliation |
Institute Ruder Boskovi?, Division of Molecular Medicine, Bijenicka 54, 10000 Zagreb, Croatia.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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