Studies on Biosynthesis of a Type II Sex Pheromone, Transportation of the Hydrophobic Precursor in a Body, and Mechanism of Their Hormonal Regulation

日本尺蠖（Ascotisselenariacretacea，Assc）的处女雌虫分泌C_193，4-环氧-6，9-二烯吸引雄虫。先前的实验表明，环氧烯基信息素是从信息素腺体外产生的相应的3，6，9-三烯生物合成的，并在与脂蛋白结合后通过血淋巴转运到信息素腺体。为了阐明这种转运，在SDS-PAGE上显示两条带（apoLp I与约250 kDa和apoLp II与约80 kDa）的雌蛾中的高密度脂蛋白（HDLp）通过KBr平衡密度梯度超离心纯化，并且通过从分离的蛋白质的溶剂提取物的GC-MS分析证实三烯的缔合。接下来，通过将氘代三烯基前体局部应用于雌性的睾丸来揭示HDL p的作用。三烯基前体与HDLp相关。在它们的信息素腺体中，三烯和氘代环氧信息素， 关于我们 重新检测，表明三烯通过血淋巴的运动。对白垩纪A.s.的雄蛾和白垩纪A.s.的雌蛾进行的实验表明，HDLp与局部施用的三烯具有相同的关联。这一结果表明，白垩盲蝽雌虫体内并不发育专门运输三烯的HDLp，而是象许多鳞翅目昆虫一样，在食道下神经节（SG）中产生一种信息素生物合成激活神经肽（PBAN），以调节HDLp在信息素腺体中的生物合成。我们已经分离出Assc-PBAN cDNA，它编码181个氨基酸，包括一个PBAN同源物和四个其他推测的肽：滞育激素同源物、α-SG神经肽（SGNP）、β-SGNP和γ-SGNP，它们在C-末端都有一个FXPR（K）L基序。虽然已从15个其他物种中鉴定出30-35个氨基酸的PBAN，但Assc-PBAN同源物由28个氨基酸组成，与其他物种相比同源性较低（<46%）。具有8个氨基酸的Assc-β-SGNP也比其他β-SGNP（16-22个氨基酸）短。此外，所有已知的PBAN cDNA在β-SGNP和PBAN之间都有一个GRR序列作为切割位点，而Assc-PBAN cDNA在相应位置显示了一个不寻常的GR序列，表明β-SGNP和PBAN之间可能存在非切割。天然信息素单肽的色谱行为揭示了包括βSGNP的Assc-PBAN的独特结构，即，SVDFTPRLGRQLVDVPQRQQIEEDRLGSRTRFFSPRL-NH_2，首次从产生环氧烯基性信息素的昆虫中测定了PBAN。少

Virgin females of the Japanese giant looper (Ascotis selenaria cretacea, Assc) in the family of Geometridae secrete C_19 3,4-epoxy-6,9-diene to attract males. Previous experiments have suggested that the epoxyalkenyl pheromone is biosynthesized from the corresponding 3,6,9-triene produced outside of a pheromone gland and transported to it via hemolymph after association with lipophorin. In order to clarify this transport, high-density lipophorin (HDLp) in the female moths showing two bands (apoLp I with ca.250 kDa and apoLp II with ca.80 kDa) on an SDS-PAGE was purified by KBr equilibrium density-gradient ultracentrifugation, and the association of the triene was confirmed by GC-MS analysis of a solvent extract from the isolated protein. Next, the role of HDLp was revealed by a topical application of the deuterated trienyl precursor to the abdomens of the females. The trienyl precursor was associated with HDLp. In their pheromone glands, the triene and the deuterated epoxy pheromone we … More re detected, indicating movement of the triene via the hemolymph. Experiments with male moths of A.s.cretacea and female moths of Bombyx moil showed the same association of HDLp with the triene topically applied. This result suggested that the adult females of A.s.cretacea did not develop HDLp specialized in the triene transport.To regulate its biosynthesis in the pheromone gland, Assc females produce a pheromone biosynthesis-activating neuropeptide (PBAN) in the suboesophageal ganglion (SG), as do females in many lepidopteran species. We have isolated Assc-PBAN cDNA, which encodes 181 amino acids, including a PBAN homologue and four other putative peptides : a diapause hormone homologue, α-SG neuropeptide (SGNP), β-SGNP, and γ-SGNP, all of which shared an FXPR(K)L motif on their C-termini. Although PBANs with 30-35 amino acids have been characterized from 15 other species, the Assc-PBAN homologue consisted of 28 amino acids and showed low homology (< 46 %) compared with the others. Assc-β-SGNP with 8 amino acids was also shorter than the other β-SGNPs (16-22 amino acids). Furthermore, all of the known PBAN cDNAs have a GRR sequence between β-SGNP and PBAN as a cleavage site, but the Assc-PBAN cDNA showed an unusual GR sequence at the corresponding position, indicating the possibility of non-cleavage between the β-SGNP and PBAN. The chromatographic behavior of the natural pheromonotoropic peptide revealed the unique structure of Assc-PBAN including βSGNP, i.e., SVDFTPRLGRQLVDDVPQRQQIEEDRLGSRTRFFSPRL-NH_2, as the first determination of PBAN from the insects producing an epoxyalkenyl sex pheromone. Less