| Title: | Ipsdienol dehydrogenase (IDOLDH): a novel oxidoreductase important for Ips pini pheromone production |
| Author(s): | Figueroa-Teran R; Welch WH; Blomquist GJ; Tittiger C; |
| Address: | "Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV 89557, United States" |
| DOI: | 10.1016/j.ibmb.2011.10.009 |
| ISSN/ISBN: | 1879-0240 (Electronic) 0965-1748 (Linking) |
| Abstract: | "Ipsdienone (2-methyl-6-methylene-2,7-octadien-4-one) is an important intermediate in the biosynthesis of pheromonal ipsdienol (2-methyl-6-methylene-2,7-octadien-4-ol) and ipsenol (2-methyl-6-methylene-7-octen-4-ol) in male pine engraver beetles, Ips pini (Say). A novel ipsdienol dehydrogenase (IDOLDH) with a pheromone-biosynthetic gene expression pattern was cloned, expressed, functionally characterized, and its cellular localization analyzed. The cDNA has a 762nt ORF encoding a 253 amino acid predicted translation product of 28kDa and pI 5.8. The protein has conserved motifs of the Cp2 subfamily of 'classical' short-chain dehydrogenases. Transcript levels were highest in pheromone producing tissue: the anterior midgut of fed males. The protein was detected only in male midguts and localized in the cytosolic fraction of midgut cells. Recombinant IDOLDH was produced in Sf9 cells using a baculovirus expression system. Enzyme assays of protein preparations showed IDOLDH used both NAD(+) and NADP(+) as coenzymes with specific activities in the nanomole range. Enzyme assays and GC/MS analysis showed that IDOLDH catalyzed the oxidation of racemic ipsdienol and (4R)-(-)-ipsdienol to form ipsdienone, while (4S)-(+)-ipsdienol was not a substrate. These data strongly implicate IDOLDH as an enzyme involved in terminal pheromone-biosynthetic steps, likely functioning to 'tune' ipsdienol enantiomeric ratios" |
| Keywords: | "Acyclic Monoterpenes Amino Acid Sequence Animals Base Sequence Cell Line Cloning, Molecular Coleoptera/*enzymology Female Insect Proteins/*metabolism Male Molecular Sequence Data Monoterpenes/*metabolism Octanols/*metabolism Oxidoreductases/*metabolism Ph;" |
| Notes: | "MedlineFigueroa-Teran, Rubi Welch, William H Blomquist, Gary J Tittiger, Claus eng Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. England 2011/11/22 Insect Biochem Mol Biol. 2012 Feb; 42(2):81-90. doi: 10.1016/j.ibmb.2011.10.009. Epub 2011 Nov 11" |
