Yeast peroxisomal multifunctional enzyme: (3R)-hydroxyacyl-CoA dehydrogenase domains A and B are required for optimal growth on oleic acid
| Autor: | Tuomo Glumoff, Kirsi M. Siivari, Mari S. Marttila, J. Kalervo Hiltunen, Yong-Mei Qin, Antti M. Haapalainen |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Saccharomyces cerevisiae
Molecular Sequence Data Dehydrogenase Biochemistry Serine Candida tropicalis Peroxisomes Humans Amino Acid Sequence Molecular Biology Alcohol dehydrogenase Candida DNA Primers chemistry.chemical_classification biology Base Sequence Sequence Homology Amino Acid 3-Hydroxyacyl CoA Dehydrogenases Cell Biology Peroxisome biology.organism_classification Molecular biology Yeast Recombinant Proteins Kinetics Enzyme chemistry biology.protein Chromatography Gel Oxidation-Reduction Oleic Acid |
| Zdroj: | The Journal of biological chemistry. 274(40) |
| ISSN: | 0021-9258 |
| Popis: | The yeast peroxisomal (3R)-hydroxyacyl-CoA dehydrogenase/2-enoyl-CoA hydratase 2 (multifunctional enzyme type 2; MFE-2) has two N-terminal domains belonging to the short chain alcohol dehydrogenase/reductase superfamily. To investigate the physiological roles of these domains, here called A and B, Saccharomyces cerevisiae fox-2 cells (devoid of Sc MFE-2) were taken as a model system. Gly(16) and Gly(329) of the S. cerevisiae A and B domains, corresponding to Gly(16), which is mutated in the human MFE-2 deficiency, were mutated to serine and cloned into the yeast expression plasmid pYE352. In oleic acid medium, fox-2 cells transformed with pYE352:: ScMFE-2(aDelta) and pYE352::ScMFE-2(bDelta) grew slower than cells transformed with pYE352::ScMFE-2, whereas cells transformed with pYE352::ScMFE-2(aDeltabDelta) failed to grow. Candida tropicalis MFE-2 with a deleted hydratase 2 domain (Ct MFE- 2(h2Delta)) and mutational variants of the A and B domains (Ct MFE- 2(h2DeltaaDelta), Ct MFE- 2(h2DeltabDelta), and Ct MFE- 2(h2DeltaaDeltabDelta)) were overexpressed and characterized. All proteins were dimers with similar secondary structure elements. Both wild type domains were enzymatically active, with the B domain showing the highest activity with short chain and the A domain with medium and long chain (3R)-hydroxyacyl-CoA substrates. The data show that the dehydrogenase domains of yeast MFE-2 have different substrate specificities required to allow the yeast to propagate optimally on fatty acids as the carbon source. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|