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G K Chambers

Publications and source records attributed to G K Chambers.

At least 55 records · Page 3Linked to original sources
PubMed1984

The purification and biochemical properties of alcohol dehydrogenase--"fast (Chateau Douglas)" from Drosophila melanogaster.

Alcohol dehydrogenase has been purified from Drosophila melanogaster lines bearing the AdhF, AdhS, and AdhFCh.D. alleles. Biochemical investigations show that the properties of the purified enzymes are very similar to those of crude enzyme extracts except that the pure enzymes are more heat stable. ADH-FCh.D. resembles ADH-S very closely in specific activity, substrate specificity, and a number of kinetic parameters including limiting values for Km(app.) for ethanol. However, it is considerably more heat stable than either of the two common variants. ADH-F differs from ADH-S and ADH-FCh.D. particularly with regard to the rate of oxidation of secondary alcohols. Atomic absorbtion spectroscopy shows that all three allozymes lack zinc or other divalent cations as active-site components. Peptide mapping experiments identify one very active cysteinyl residue; and amide residues in the NAD+ binding domain.

Alcohol Dehydrogenase↗
PubMed1984

Variation in the biochemical properties of the Drosophila alcohol dehydrogenase allozymes.

Thirteen Drosophila Adh variants have been characterized with respect to gene expression, substrate preference, thermostability, and specific activity. The results suggest that the variants may be grouped into two biochemical classes, typified by the properties of the two most common enzyme forms, ADH-F and ADH-S. Membership of these classes cannot be predicted from electrophoretic mobility, nor is any simple classification possible with regard to the characteristics of level of gene expression (in terms of ADH activity or ADH protein) or thermostability of the gene product.

Alcohol Dehydrogenase↗
PubMed1981

Latitudinal relationships of esterase-6 and phosphoglucomutase gene frequencies in Drosophila melanogaster.

Geographic variation in Esterase-6 (Est-6) and Phosphoglucomutase (Pgm) gene frequencies in Australasian populations of Drosophila melanogaster are compared with analogous data collated from 16 previous reports for North America and Europe/Asia. A large-scale latitudinal cline is found on all three zoogeographic zones for Est-6 and overall, Est-61.00 frequency increases from about 20 per cent around 20 degrees latitude to about 80 per cent approaching 50 degrees latitude. In contrast, there is no consistent evidence for the latitudinal cline in Pgm gene frequencies in any of the three zones with Pgm1.00 frequency generally about 85 per cent and Pgm1.20 and Pgm0.70 frequencies each between 5 per cent and 10 per cent. The consistent Est-6 clines are attributed to latitudinal selection gradients but not consistent correlations are found between Est-6 gene frequencies and maximum or minimum temperature or rainfall which might be associated with these gradients. The directions of the Est-6 clines in fact run counter to expectations based on the in vitro thermostabilities of the respective allozymes.

Animals↗
PubMed1981

An electrophoretically cryptic alcohol dehydrogenase variant in Drosophila melanogaster. III. Biochemical properties and comparison with common enzyme forms.

The biochemical properties of the heat-stable alcohol dehydrogenase variant ADH-FCh.D. have been investigated and compared with those of the two common enzyme forms ADH-F and ADH-S. The results show that ADH-F and ADH-S differ with respect to substrate specificity, their response to high concentrations of secondary alcohols and their apparent Michaelis constants for three alcohols in two different buffer systems. In all these tests the enzyme ADH-FCh.D. resembles ADH-S much more closely than ADH-F. It is concluded that if natural selection is to distinguish between the alleles AdhS and AdhFCh.D. then it most probably does so on the basis of the superior thermostability of ADH-FCh.D. The biochemical properties of all three enzymes are discussed in relation to the role of alcohol dehydrogenase in the exploitation of alcohol by D. melanogaster.

Alcohol Dehydrogenase↗
PubMed1981

Structural analysis of an electrophoretically cryptic alcohol dehydrogenase variant from an Australian population of Drosophila melanogaster.

The existence of geographically widespread clines in genetic polymorphisms is persuasive evidence that the distribution of such genetic variance is determined by natural selection. However, when comparing clines it is important to be certain that identical structural genes are involved. We report a structural difference (proline-214 to serine) between the product of AdhF and an electrophoretically cryptic heat-stable variant isolated from an Australian natural population, ADH-FCh.D. ("fast" Chateau Douglas). The biochemical properties of this new variant must be taken into consideration when attempting to formulate a causal explanation of the maintenance of the three identified Adh alleles. Our data also show that the products of an AdhF and an AdhS allele in Drosophila melanogaster in an Australian population are identical over 70% of their amino acid sequences with their North American counterparts.

Alcohol Oxidoreductases↗
PubMed1980

An electrophoretically cryptic alcohol dehydrogenase variant in Drosophila melanogaster. I. Activity ratios, thermostability, genetic localization and comparison with two other thermostable variants.

The alcohol dehydrogenase (ADH) variant ADH-FCh.D. has a secondary alcohol/primary alcohol activity ratio characteristic of ADH-S although it has an electrophoretic mobility inseparable from ADH-F. ADH-FCh.D. is distinguished from these two common ADH variants by being much more thermostable. Genetic analysis suggests tht ADH-FCh.D. is specified by an allele at the Adh locus. Biochemical comparisons show that ADH-FCh.D. has the same electrophoretic mobility, activity ratio and thermostability as the two other heat-resistant variants which have been reported, ADH-F71K in Europe and ADH-Fr in North America. The geographically widespread distribution of a thermostable ADH variant within the ADH-F electrophoretic class indicates that it should be considered in attempts to explain the Adh polymorphism in natural populations.

Alcohol Oxidoreductases↗
PubMed1978

Alcohol-oxidizing enzymes in 13 Drosophila species.

Starch and polyacrylamide gel electrophoresis were used to ascertain the substrate specificities of alcohol-oxidizing enzymes in 13 Drosophila species. The substrates used were a variety of long- and short-chain aliphatic alcohols, one aromatic alcohol, and benzaldehyde. Only one enzyme (product of a single-gene locus) showed significant NAD+-dependent alcohol dehydrogenase activity with short-chain aliphatic alcohols. The 13 species, belonging to four different Drosophila groups, all showed a similar complement of alcohol-oxidizing enzymes, although differences in electrophoretic mobility and in levels of activity existed from species to species. These findings are relevant to the adaptation of Drosophila to alcohol environments.

Alcohol Oxidoreductases↗
PubMed1978

Structural analysis of the ADHS electromorph of Drosophila melanogaster.

Population geneticists have often determined the fitness differences that account for the dynamics of naturally occurring genetic polymorphisms. However, to understand causal aspects of evolutionary processes requires, in addition, investigation of the physiological and molecular structural differences underlying adaptively significant genetic polymorphisms. The characteristics of the alcohol dehydrogenase gene--enzyme system in Drosophila melanogaster make it well suited for this kind of study. Natural populations of this species are polymorphic for two electrophoretically detectable variants, ADHF and ADHS, of the enzyme. Structural studies reported here reveal that the two variants differ by (at least) a single amino acid replacement, threonine in ADHF for lysine in ADHS.

Alcohol Oxidoreductases↗
PubMed1977

Adaptive response due to changes in gene regulation: a study with Drosophila.

In spite of the critical role of the process of adaptation in evolution, there are few detailed studies of the genotypic and molecular basis of the process. Drosophila melanogaster flies selected for increased tolerance to ethanol exhibited higher levels of alcohol dehydrogenase (alcohol:NAD+ oxidoreductase; EC 1.1.1.1) activity than unselected controls. A series of tests (electrophoresis, product inhibition, temperature stability, pH optima, substrate specificity, and Michaelis constants) gave no evidence of structural differences in the enzyme of the selected and the control flies. However, quantitative immunological assays showed that the selected flies contained significantly higher amounts of alcohol dehydrogenase. Adaptation of the selected flies to higher alcohol tolerance has most likely taken place by changes not in the structural gene locus coding for the enzyme, but by regulatory changes affecting the amount of gene product.

Adaptation, Biological↗
PubMed1975

The amino acid sequence of Neurospora NADP-specific glutamate dehydrogenase. The tryptic peptides.

The NADP-specific glutamate dehydrogenase of Neurospora crassa was digested with trypsin, and peptides accounting for 441 out of the 452 residues of the polypeptide chain were isolated and substantially sequenced. Additional experimental detail has been deposited as Supplementary Publication SUP 50052 (11 pages) with the British Library (Lending Division), Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained under the terms given in Biochem J. (1975) 145, 5.

Amides↗
PubMed1975

The amino acid sequence of Neurospora NADP-specific glutamate dehydrogenase. Peptic and chymotryptic peptides and the complete sequence.

Peptic and chymotryptic peptides were isolated form the NADP-specific glutamate dehydrogenase of Neurospora crassa and substantially sequenced. Out of 452 residues in the polypeptide chain, 265 were recovered in the peptic and 427 in the chymotryptic peptides. Together with the tryptic peptides [Wootton, J. C., Taylor, J. G., Jackson, A. A., Chambers, G. K. & Fincham, J. R. S. (1975) Biochem. J. 149, 749-755], these establish the complete sequence of the chain, including the acid and amide assignments, except for seven places where overlaps are inadequate. These remaining alignments are deduced from information on the CNBr fragments obtained in another laboratory [Blumenthal, K. M., Moon, K. & Smith, E. L. (1975), J. Biol. Chem. 250, 3644-3654]. Further information has been deposited as Supplementary Publication SUP 50054 (17 pages) with the British Library (Lending Division), Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained under the terms given in Biochem. J. (1975) 145, 5.

Amides↗
PubMed1974

Amino-acid sequence of NADP-specific glutamate dehydrogenase of neurospora crassa.

A tentative primary structure of the NADP-specific glutamate dehydrogenase [L-glutamate: NADP oxidoreductase (deaminating), EC 1.4.1.4] from Neurospora crassa has been determined. The proposed sequence contains 452 amino-acid residues in each of the identical subunits of the hexameric enzyme. Comparison of the sequence with that of the bovine liver enzyme reveals considerable homology in the amino-terminal portion of the chain, including the vicinity of the reactive lysine, with only shorter stretches of homology within the carboxyl-terminal regions. The significance of this distribution of homologous regions is discussed.

Amino Acid Sequence↗