A novel terpene synthase controls differences in anti-aphrodisiac pheromone production between closely related Heliconius butterflies

Plants and insects often use the same compounds for chemical communication, but not much is known about the genetics of convergent evolution of chemical signals. The terpene (E)-β-ocimene is a common component of floral scent and is also used by the butterfly Heliconius melpomene as an anti-aphrodisiac pheromone. While the biosynthesis of terpenes has been described in plants and microorganisms, few terpene synthases (TPSs) have been identified in insects. Here, we study the recent divergence of 2 species, H. melpomene and Heliconius cydno, which differ in the presence of (E)-β-ocimene; combining linkage mapping, gene expression, and functional analyses, we identify 2 novel TPSs. Furthermore, we demonstrate that one, HmelOS, is able to synthesise (E)-β-ocimene in vitro. We find no evidence for TPS activity in HcydOS (HmelOS ortholog of H. cydno), suggesting that the loss of (E)-β-ocimene in this species is the result of coding, not regulatory, differences. The TPS enzymes we discovered are unrelated to previously described plant and insect TPSs, demonstrating that chemical convergence has independent evolutionary origins.

Plants and insects often use the same compounds for chemical communication, but little is known about the convergent evolution of such chemical signals. This study identifies a novel terpene synthase involved in production of an anti-aphrodisiac pheromone by the butterfly Heliconius melpomene. This enzyme is unrelated to other insect terpene synthases, providing evidence that the ability to synthesise terpenes has arisen multiple times independently within the insects.     

A large photoreactive particle from Chromatium vinosum chromatophores

Large photoreactive particles from Chromatium vinosum are obtained pure and in high yield by using a mixture of detergents at high ionic strength to dissociate the chromatophore membrane. The particles contain all of the secondary electron acceptor of the chromatophores and about half of the cytochrome. Their content of ubiquinone is greatly enriched as compared with chromatophores. The individual particles have an estimated molecular weight of between 650 000 and 810 000.Gel electrophoresis of the preparation in sodium dodecylsulfate shows polypeptides with molecular weights of 50–45 000, 30 000, 27 000, 22 000 and 12 000. The 50–45 000 components are cytochromes. The 30 000, 27 000 and 22 000 components may be analogous to the triad of polypeptides present in Rhodopseudomonas spheroides reaction centers. The non-cytochrome components are partly soluble in chloroform/methanol.Aggregates of particles appear in these preparations. Electron microscopy of the aggregates demonstrates rectilinear lattices of isodiametric particles, 120 Å in diameter. These sheet-like structures are one unit thick and typically contain 9–16 members. They appear to arise by aggregation during isolation but are probably similar to native aggregates apparent within chromatophores after treatment with detergents at low salt concentration.     

On the mechanism of action of methyl chymotrypsin

The properties of a-chymotrypsin methylated at histidine-57 were examined to explain the mechanism of this enzyme which is about 10⁵ times less active than chymotrypsin. Studies on the protein showed (i) an alteration in the acyl and leaving group specificity, (ii) decreased binding of some protein protease inhibitors by methyl chymotrypsin, (iii) lack of dimerization of methyl chymotrypsin at low pH, (iv) decreased stability of methyl chymotrypsin in urea, (v) a larger solvent deuterium isotope effect with methyl chymotrypsin, and (vi) decreased binding of a tetrahedral intermediate analog to methyl chymotrypsin. These properties suggest that while only subtle alterations occur in the active site upon methylation of His-57, the transition state and the tetrahedral intermediate are destabilized but not to the same extent. General base catalysis remains an integral feature of the hydrolytic mechanism of the modified chymotrypsin, and the base appears to be the methylated nitrogen of the imidazole moiety of His-57.