Year-end Sale % OFF 
Abstract
Males of the parasitic wasp genus Nasonia use blends of chiral hydroxylactones as sex pheromones to attract conspecific females. Whereas all Nasonia species use a mixture of (4R,5S)-5-hydroxy-4-decanolide (RS) and 4-methylquinazoline (MQ) as sex pheromones, Nasonia vitripennis evolved (4R,5R)-5-hydroxy-4-decanolide (RR) as an extra sex pheromone component. We recently identified and functionally characterized three short-chain dehydrogenases/reductases (SDRs) NV10127, NV10128, and NV10129 that are capable of catalyzing the epimerization of RS to RR via (4R)-5-oxo-4-decanolide (ODL) as intermediate. Despite their very high sequence identities of 88–98%, these proteins differ drastically in their ability to epimerize RS to RR and in their stereoselectivity when reducing ODL to RR/RS. Here, in order to unravel the sequence differences underlying these varying functional properties of NV1027, NV10128 and NV10129, we created chimeras of the three enzymes and monitored their catalytic activities in vitro. The results show that a few amino acid changes at the C-termini and active sites of Nasonia vitripennis SDRs lead to substantially altered RS to RR epimerization and ODL-reduction activities. Thus, our study adds to the understanding of pheromone evolution by showing that subtle mutations in key biosynthetic enzymes can result in drastic effects on the composition of chemical signals.
Highlights
Sex pheromones are pivotal for mate location, recognition, and acceptance in insects[1,2]
By comparing the amino acid sequences of NV10127 and NV10128 we found a total of 50 amino acid differences (30 non-conservative and 20 conservative) along with a short gap in NV10128
The NV10127-29 enzymes investigated in this study have sequence identities of 88–98% and the data obtained with the chimeras confirm that the evolution of new functional properties can be elicited by very small changes in the coding sequence of N. vitripennis short-chain dehydrogenases/reductases (SDRs)
Summary
Sex pheromones are pivotal for mate location, recognition, and acceptance in insects[1,2]. NV10127, the most abundant enzyme in male Nv pheromone glands, was drastically less efficient at epimerizing RS to RR and was capable of stoichiometrically reducing ODL in favor of the RR product. As a first step to investigate the structure-activity relationship underlying Nv SDRs activity, we aimed to identify sequence motifs for the different functional properties of NV10127, NV10128, and NV10129. To this end, we used site-directed mutagenesis to generate chimeras of the three enzymes, heterologously expressed the corresponding genes in Escherichia coli, purified the proteins to homogeneity and monitored their catalytic activities in vitro. Our results show that subtle sequence variations of SDRs result in drastic functional consequences and provide novel insights in the sequence-related structure/activity relationships underlying the enzymatic epimerization of natural products
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
