Farnesyl biliverdins IXα are novel ligands of biliproteins from moths of the Noctuoidea superfamily: A chemosystematic view of the Lepidoptera.

Bilins, derived from biliverdin IXα, are known from animals, plants and microorganisms, where they play vital roles as light-absorbing pigments. Bilins occur also in many insects. Recently, we discovered in insects a novel structural type of bilins with a farnesyl substituent at pyrrole ring A of biliverdin IXα.

cDNA sequences of two arylphorin subunits of an insect biliprotein: phylogenetic differences and gene duplications during evolution of hexamerins-implications for hexamer formation.

Arylphorins represent a conserved class of hexameric ∼500 kDa insect hemolymph glycoproteins, rich in aromatic amino acids, which are produced in large quantities at the larval stage as reserves for metamorphosis and egg development. The recently isolated arylphorin from the moth Cerura vinula is unique in being complexed to a novel farnesylated bilin. Protein sequencing suggested the presence of two different ∼85 kDa subunits.

Binding of imidacloprid, thiamethoxam and N-desmethylthiamethoxam to nicotinic receptors of Myzus persicae: pharmacological profiling using neonicotinoids, natural agonists and antagonists.

Background: The increasing structural diversity of the neonicotinoid class of insecticides presently used in crop protection calls for a more detailed analysis of their mode of action at their cellular targets, the nicotinic acetylcholine receptors.

Results: Comparative radioligand binding studies using membranes of Myzus persicae (Sulzer) and representatives of the chloropyridyl subclass (imidacloprid), the chlorothiazolyl subclass (thiamethoxam), the tetrahydrofuranyl subclass (dinotefuran), as well as the novel sulfoximine type (sulfoxaflor), which is not a neonicotinoid, reveal significant differences in the number of binding sites, the displacing potencies and the mode of binding interference. Furthermore, the mode of interaction of [ H]thiamethoxam and the nicotinic antagonists methyllycaconitine and dihydro-β-erythroidine is unique, with Hill values of >1, clearly different to the values of around unity for [ H]imidacloprid and [ H]N-desmethylthiamethoxam.

Very high-density lipoprotein and vitellin as carriers of novel biliverdins IXα with a farnesyl side-chain presumably derived from heme A in Spodoptera littoralis.

Bilins in complex with specific proteins play key roles in many forms of life. Biliproteins have also been isolated from insects; however, structural details are rare and possible functions largely unknown. Recently, we identified a high-molecular weight biliprotein from a moth, Cerura vinula, as an arylphorin-type hexameric storage protein linked to a novel farnesyl biliverdin IXα; its unusual structure suggests formation by cleavage of mitochondrial heme A.

Structure of a novel farnesylated bilin from an insect--formation by α-cleavage of heme A of mitochondrial cytochrome c oxidases?

Biliproteins are present in almost all forms of life, and many of them play vital roles in photobiology. The bilin ligand of a recently characterized 500-kDa biliprotein from an insect has been isolated and its structure elucidated with chemical and spectroscopic techniques (UV-visible, IR, MS, NMR, and CD). This blue pigment, named CV-bilin, represents a unique high molecular mass derivative of biliverdin IXα, with an unusual 10E-configuration and a molecular mass of 852 Da, corresponding to C48H60N4O10.

C-26 vs. C-27 hydroxylation of insect steroid hormones: regioselectivity of a microsomal cytochrome P450 from a hormone-resistant cell line.

Hydroxylation of steroids at one of the side chain terminal methyl groups, commonly linked to C-26, represents an important regulatory step established in many phyla. Discrimination between the two sites, C-26 and C-27, requires knowing the stereochemistry of the products. 26-Hydroxylation of the insect steroid hormone 20-hydroxyecdysone by a microsomal cytochrome P450 was previously found to be responsible for hormonal resistance in a Chironomus cell line mainly producing the (25S)-epimer of 20,26-dihydroxyecdysone.

Isolation, characterisation and molecular imaging of a high-molecular-weight insect biliprotein, a member of the hexameric arylphorin protein family.

The abundant blue hemolymph protein of the last instar larvae of the moth Cerura vinula was purified and characterized by protein-analytical, spectroscopic and electron microscopic methods. Amino acid sequences obtained from a large number of cleavage peptides revealed a high level of similarity of the blue protein with arylphorins from a number of other moth species. In particular, there is a high abundance of the aromatic amino acids tyrosine and phenylalanine amounting to about 19% of total amino acids and a low content of methionine (0.

Porphobilinogen synthase from the butterfly, Pieris brassicae: purification and comparative characterization.

Porphobilinogen represents a key building block of tetrapyrroles serving as functional ligands of many vitally important proteins. Here we report the first purification of porphobilinogen synthase (PBGS) from whole insects by sequentially employing two modes of native electrophoresis on polyacrylamide gels subsequent to more conventional procedures. Using adults of Pieris brassicae L.

Stable free radicals in insect cuticles: electron spin resonance spectroscopy reveals differences between melanization and sclerotization.

Insect cuticles (exuviae; cast skins) were examined for the first time by ESR spectroscopy for the presence of stable free radicals, as found in melanins. All cuticles, except those from a locust albino strain, irrespective of the presence of melanin, provided single-line signals of varied g-values and linewidths. The ESR signals of melanins, isolated or in cuticles, were characterized by g-values <2.

Butterfly wings, a new site of porphyrin synthesis and cleavage: studies on the expression of the lipocalin bilin-binding protein in Pieris brassicae.

The bilin-binding protein (BBP), a member of the lipocalin protein superfamily, is synthesized mainly in last instar larvae and in late pupae and newly emerged adults of Pieris brassicae, as previously reported. Here we present results from Northern blot analysis of the BBP gene transcript and from in vitro studies of holo-BBP biosynthesis with isolated wings using [14C]5-aminolevulinic acid as a precursor to the bilin ligand, [14C]-amino acids to label the apo-protein and inhibitors for both processes. Our combined data clearly demonstrate that BBP, which accumulates around pupa-adult transformation, is produced as holoprotein in the developing wings, while the BBP gene transcript is no longer detected in the rest of the body.

The insecticide pymetrozine selectively affects chordotonal mechanoreceptors.

Pymetrozine is a neuroactive insecticide but its site of action in the nervous system is unknown. Based on previous studies of symptoms in the locust, the feedback loop controlling the femur-tibia joint of the middle leg was chosen to examine possible targets of the insecticide. The femoral chordotonal organ, which monitors joint position and movement, turned out to be the primary site of pymetrozine action, while interneurons, motoneurons and central motor control circuitry in general did not noticeably respond to the insecticide.

Developmental profiles of 5-aminolevulinate, porphobilinogen and porphobilinogen synthase activity in Pieris brassicae related to the synthesis of the bilin-binding protein.

The bilin-binding protein (BBP) occurs as a major soluble protein in haemolymph, fat body, epidermis and wings of Pieris brassicae. It is a member of the lipocalin protein superfamily with yet unknown function. Here, we studied the developmental regulation of tetrapyrrole biosynthesis that provides the bilin ligand as the predominating end product.

The serotonergic system is involved in feeding inhibition by pymetrozine. Comparative studies on a locust (Locusta migratoria) and an aphid (Myzus persicae).

Pymetrozine inhibits feeding in aphids immediately after application without producing visible neurotoxic effects, as previously reported. In the present work, Locusta migratoria, though not a plant-sucking insect, was found to respond to pymetrozine by displaying unique symptoms, which were lifting and stretching of the hindlegs, in addition to inhibition of feeding. In locust, pymetrozine enhanced spontaneous spike discharge of the metathoracic and suboesophageal ganglia in situ at nanomolar concentrations.

Comparative analysis of neonicotinoid binding to insect membranes: II. An unusual high affinity site for [3H]thiamethoxam in Myzus persicae and Aphis craccivora.

Neonicotinoids represent a class of insect-selective ligands of nicotinic acetylcholine receptors. Imidacloprid, the first commercially used neonicotinoid insecticide, has been studied on neuronal preparations from many insects to date. Here we report first intrinsic binding data of thiamethoxam, using membranes from Myzus persicae Sulzer and Aphis craccivora Koch.

Comparative analysis of neonicotinoid binding to insect membranes: I. A structure-activity study of the mode of [3H]imidacloprid displacement in Myzus persicae and Aphis craccivora.

Neonicotinoids bind selectively to insect nicotinic acetylcholine receptors with nanomolar affinity to act as potent insecticides. While the members of the neonicotinoid class have many structural features in common, it is not known whether they also share the same mode of binding to the target receptor. Previous competition studies with [3H]imidacloprid, the first commercialised neonicotinoid, indicated that thiamethoxam, representing a novel structural sub-class, may bind in a different way from that of other neonicotinoids.

Diastereomeric ecdysteroids with a cyclic hemiacetal in the side chain produced by cytochrome P450 in hormonally resistant insect cells.

A microsomal cytochrome P450 from a cell line of the insect Chironomus tentans has been shown to hydroxylate the steroid hormone 20-hydroxyecdysone at C(26) to yield 20,26-dihydroxyecdysone, P1, which is further metabolized to P2 and P3. Based on (1)H NMR studies, acetonide formation and quantum chemical calculations, P2 and P3 represent novel slowly interconvertible geometrical isomers, occurring at a 3:1 ratio, presumably arising from hemiacetal formation between the 26-aldehyde group and the 22R-hydroxyl group to build a tetrahydropyran ring in the side chain. The stereochemistry at C(26) was S in P2 (trans-diol) and R in P3 (cis-diol), respectively.