Identification and characterisation of two allelic forms of human alcohol dehydrogenase 2

The human alcohol dehydrogenase system is comprised of multiple forms that catalyse the oxidation/reduction of a large variety of alcohols and aldehydes. A transition that results in an Ile308Val substitution was identified in the human ADH2 gene by single-strand conformation polymorphism analysis....

Full description

Saved in:
Bibliographic Details
Published in:Cellular and molecular life sciences : CMLS 2002-03, Vol.59 (3), p.552-559
Main Authors: Strömberg, P, Svensson, S, Hedberg, J J, Nordling, E, Höög, J O
Format: Article
Language:English
Subjects:
Alcohol
Alcohol Dehydrogenase - chemistry
Alcohol Dehydrogenase - genetics
Alcohol Dehydrogenase - metabolism
Alcohols
Aldehydes
Alleles
Amino Acid Sequence
Amino Acid Substitution - genetics
Base Sequence
Binding Sites
Dehydrogenase
E coli
Enzyme Stability
Enzymes
Ethanol
Exons - genetics
Gene Frequency
Genetic diversity
Humans
Kinetics
Models, Molecular
Molecular biology
Mutagenesis
Polymerase Chain Reaction
Polymorphism
Polymorphism, Genetic - genetics
Protein Conformation
Proteins
Structure-Activity Relationship
Substrate Specificity
Temperature
Time Factors
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The human alcohol dehydrogenase system is comprised of multiple forms that catalyse the oxidation/reduction of a large variety of alcohols and aldehydes. A transition that results in an Ile308Val substitution was identified in the human ADH2 gene by single-strand conformation polymorphism analysis. Screening a Swedish population revealed that Val308 was the most frequent allele (73%), and site-directed mutagenesis was used to obtain both allelozymes, which were expressed in Escherichia coli for characterisation. Thermostability was assayed by activity measurements and circular dichroism spectroscopy. The results showed that the 308Val substitution decreases protein stability, as compared to the Ile308 variant, an effect also demonstrated during prolonged storage. Ethanol, octanol, 12-hydroxydodecanoic acid and all-trans retinol were used as model substrates and, generally, slightly higher Km values were observed with Val at position 308. Finally, homology modelling, from mouse ADH2, further supported the decreased stability of the Val308 variant and located position 308 in the subunit interface of the molecule and in the vicinity of the active-site pocket entrance. In conclusion, the Ile308Val substitution represents a novel functional polymorphism within the human alcohol dehydrogenase gene cluster that may affect the metabolism of ethanol and other substrates.
ISSN:1420-682X
1420-9071
DOI:10.1007/s00018-002-8447-1