|School or College
|College of Science
|Poulter, Charles Dale
|Anderson, Matt S.; Muehlbacher, Manfred; Street, Ian P.; Proffitt, John
|Isopentenyl diphosphate:dimethylallyl diphosphate isomerase. An improved purification of the enzyme and isolation of the gene from Saccharomyces cerevisiae.
|Isopentenyl diphosphate:dimethylallyl diphosphate isomerase (IPP isomerase) is an enzyme in the isoprenoid biosynthetic pathway which catalyzes the interconversion of the primary five-carbon homoallylic and allylic diphosphate building blocks. We report a substantially improved procedure for purification of this enzyme from Saccharomyces cerevisiae. An amino-terminal sequence (35 amino acids) was obtained from a highly purified preparation of IPP isomerase. Oligonucleotide probes based on the protein sequence were used to isolate the structural gene encoding IPP isomerase from a yeast lambda library. The cloned gene encodes a 33,350-dalton polypeptide of 288 amino acids. A 3.5-kilobase EcoRI fragment containing the gene was subcloned into the yeast shuttle vector YRp17. Upon transformation with plasmids containing the insert, a 5-6-fold increase in IPP isomerase activity was seen in transformed cells relative to YRp17 controls, confirming the identity of the cloned gene. This is the first reported isolation of the gene for IPP isomerase.
|American Society for Biochemistry and Molecular Biology (ASBMB)
|Amino Acid Sequence; Base Sequence; Chromatography, Gel; Chromatography, Ion Exchange
|Carbon-Carbon Double Bond Isomerases; DNA, Fungal; Genes, Fungal; Saccharomyces cerevisiae
|J Biol Chem. 1989 Nov 15;264(32):19169-75. Anderson MS, Muehlbacher M, Street IP, Proffitt J, Poulter CD. Isopentenyl diphosphate:dimethylallyl diphosphate isomerase. An improved purification of the enzyme and isolation of the gene from Saccharomyces cerevisiae. Retieved April 16,2007 from http://www.jbc.org/cgi/reprint/264/32/19169.
|Copyright © 1989 American Society for Biochemistry and Molecular Biology. All rights reserved.