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Rapid screening of cytochromes of respiratory mutants of Saccharomyces cerevisiae. Application to the selection of strains containing novel forms of cytochrome-c oxidase
- Source :
- European Journal of Biochemistry. 213:137-145
- Publication Year :
- 1993
- Publisher :
- Wiley, 1993.
-
Abstract
- A technique has been developed for the direct analysis by visible spectrophotometry of yeast spots growing on agar plates. This allows rapid semi-quantitative estimations of cytochromes c, b and oxidase and permits the identification of strains with impaired respiratory electron flow. Results of screening of 105 mutants are presented. There appears to be a correlation between the exonic location of the mutation in COX1 of oxidase and the level of optically detectable enzyme. Mutations in cytochrome b of the bc1 complex also affect the level of expression of cytochrome oxidase and can cause either an increased or decreased level of expression of oxidase relative to the wild-type strain. Twelve strains selected by the rapid level-1 screening were grown as lawns on sections of an agar plate and resuspended for a second level of screening. Quantitative estimates have been made of the concentrations of cytochromes, the turnover number of cytochrome oxidase and the kinetics of recombination of carbon monoxide with oxidase after flash photolysis. This confirmed the validity of the rapid screening procedure, and we have identified several strains which contain high levels of a mutant form of cytochrome oxidase with properties worthy of further investigation.
- Subjects :
- chemistry.chemical_classification
Oxidase test
biology
Protein Conformation
Cytochrome b
Genes, Fungal
Mutant
Saccharomyces cerevisiae
biology.organism_classification
Biochemistry
Yeast
Electron Transport
Electron Transport Complex IV
Agar plate
Enzyme
chemistry
Mutation
biology.protein
Cytochromes
Cytochrome c oxidase
Subjects
Details
- ISSN :
- 14321033 and 00142956
- Volume :
- 213
- Database :
- OpenAIRE
- Journal :
- European Journal of Biochemistry
- Accession number :
- edsair.doi.dedup.....e4845da9e543fc17ababf98619d0f756
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1993.tb17743.x