Yeast model system for study of mammalian phosphodiesterases.
Facile manipulation and rapid regeneration have helped to establish the yeast Saccharomyces cerevisiae as a genetic workhorse. More recently, these simple eukaryotes have been used for the biochemical analysis of mammalian proteins. This article describes the use of a yeast expression system for both in vitro and in vivo assays of mammalian phosphodiesterase (PDE) activity using yeast cells devoid of endogenous PDEs. It also presents simple methodologies for the analysis of the pharmacological properties of mammalian PDEs and describes the use of a powerful genetic selection for mutant forms of PDEs with altered biochemical and pharmacological characteristics.
Atienza
J M
JM
University of California, Los Angeles, School of Medicine, 33-257 CHS, Los Angeles, California 90095-1737, USA.
Colicelli
J
J
D000818:Animals; D004351:Drug Resistance / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006358:Hot Temperature; D008322:Mammals; D010641:Phenotype; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000032:analysis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics
D004791:Enzyme Inhibitors; D010726:Phosphodiesterase Inhibitors; D010727:Phosphoric Diester Hydrolases