Abstract
Three alcohol dehydrogenases from Drosophila simulans, Drosophila virillis and Drosophila melanogaster adhS (which possesses an alloenzyme with slow electrophoretic mobility) were purified essentially to homogeneity. The purification procedure involves a new step of affinity chromatography, which efficiently lowers the amount of contaminants in the final preparation, producing a very stable enzyme. The purification procedure developed consists of a salmine sulphate precipitation, two CM-Sepharose CL-6B colume-chromatography steps, an affinity-chromatography step and a Sephacryl gel filtration. A minimum of 30-fold purification is obtained and the yield is not less than 34%. The isoelectric points and molar absorption coefficients were determined.
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- Day T. H., Hillier P. C., Clarke B. The relative quantities and catalytic activities of enzymes produced by alleles at the alcohol dehydrogenase locus in Drosophila melanogaster. Biochem Genet. 1974 Feb;11(2):155–165. doi: 10.1007/BF00485771. [DOI] [PubMed] [Google Scholar]
- Jacobson K. B., Pfuderer P. Interconversion of isoenzymes of Drosophila alcohol dehydrogenase. II. Physical characterization of the enzyme and its subunits. J Biol Chem. 1970 Aug 10;245(15):3938–3944. [PubMed] [Google Scholar]
- King J., Laemmli U. K. Polypeptides of the tail fibres of bacteriophage T4. J Mol Biol. 1971 Dec 28;62(3):465–477. doi: 10.1016/0022-2836(71)90148-3. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Thatcher D. R. Enzyme instability and proteolysis during the purification of an alcohol dehydrogenase from Drosophila melanogaster. Biochem J. 1977 May 1;163(2):317–323. doi: 10.1042/bj1630317. [DOI] [PMC free article] [PubMed] [Google Scholar]