Abstract
The neuropeptide mandibular organ (MO)-inhibiting hormone (MO-IH), synthesized and secreted from the X-organ-sinus-gland complex of the eyestalk, regulates the biosynthesis of the putative crustacean juvenile hormone, methyl farnesoate (MF). Using radiolabelled acetate as a precursor for isoprenoid biosynthesis, farnesoic acid (FA), farnesol, farnesal, MF and geranyl geraniol were detected in MOs cultured for 24 h. Treatment of MOs with extract of sinus gland inhibited the final step of biosynthesis of MF, catalysed by FA O-methyltransferase. Additionally, treatment of MOs with purified MO-IH exhibited a dose-dependent inhibition of this final step of MF synthesis. The extent of this inhibition was dependent on the ovary stage of the MO-donor animal, being maximal in MOs from animals in the early stages of ovarian development. Assay of FA O-methyltransferase activity, using [3H]FA in the presence of S-adenosyl-l-methionine, demonstrated that the enzyme was located in the cytosolic fraction of MOs and was inhibited by incubation of MOs with MO-IH prior to preparation of subcellular fractions. For cytosolic preparations taken from vitellogenic animals, both Vmax and Km were appreciably lower than for those taken from non-vitellogenic animals. Conversely, eyestalk ablation of early-vitellogenic animals, which removes the source of MO-IH in vivo, resulted in enhancement of the cytosolic FA O-methyltransferase activity. Although both Vmax and Km show an appreciable increase upon eyestalk ablation, the increased enzyme activity is probably reflected by the fact that Vmax/Km (an approximate indication of kcat) has increased 5-fold. The combined evidence demonstrates that MO-IH inhibits FA O-methyltransferase, the enzyme which catalyses the final step of MF biosynthesis in MOs.
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- Boivin D., Bilodeau D., Béliveau R. Immunochemical characterization of L-isoaspartyl-protein carboxyl methyltransferase from mammalian tissues. Biochem J. 1995 Aug 1;309(Pt 3):993–998. doi: 10.1042/bj3090993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Chen J. H., Hara T., Fisher M. J., Rees H. H. Immunological analysis of developmental changes in ecdysone 20-mono-oxygenase expression in the cotton leafworm, Spodoptera littoralis. Biochem J. 1994 May 1;299(Pt 3):711–717. doi: 10.1042/bj2990711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feyereisen R., Pratt G. E., Hamnett A. F. Enzymic synthesis of juvenile hormone in locust corpora allata: evidence for a microsomal cytochrome P-450 linked methyl farnesoate epoxidase. Eur J Biochem. 1981 Aug;118(2):231–238. doi: 10.1111/j.1432-1033.1981.tb06391.x. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S. Regulation of the mevalonate pathway. Nature. 1990 Feb 1;343(6257):425–430. doi: 10.1038/343425a0. [DOI] [PubMed] [Google Scholar]
- Keller R. Crustacean neuropeptides: structures, functions and comparative aspects. Experientia. 1992 May 15;48(5):439–448. doi: 10.1007/BF01928162. [DOI] [PubMed] [Google Scholar]
- Laufer H., Borst D., Baker F. C., Reuter C. C., Tsai L. W., Schooley D. A., Carrasco C., Sinkus M. Identification of a juvenile hormone-like compound in a crustacean. Science. 1987 Jan 9;235(4785):202–205. doi: 10.1126/science.235.4785.202. [DOI] [PubMed] [Google Scholar]
- McCaskill D., Croteau R. Procedures for the isolation and quantification of the intermediates of the mevalonic acid pathway. Anal Biochem. 1993 Nov 15;215(1):142–149. doi: 10.1006/abio.1993.1566. [DOI] [PubMed] [Google Scholar]
- Sagi A., Homola E., Laufer H. Methyl farnesoate in the prawn Macrobrachium rosenbergii: synthesis by the mandibular organ in vitro, and titers in the hemolymph. Comp Biochem Physiol B. 1991;99(4):879–882. doi: 10.1016/0305-0491(91)90157-9. [DOI] [PubMed] [Google Scholar]
- Sutherland T. D., Feyereisen R. Target of cockroach allatostatin in the pathway of juvenile hormone biosynthesis. Mol Cell Endocrinol. 1996 Jul 1;120(2):115–123. doi: 10.1016/0303-7207(96)03825-7. [DOI] [PubMed] [Google Scholar]
- Taketomi Y., Kawano Y. Ultrastructure of the mandibular organ of the shrimp, Penaeus japonicus, in untreated and experimentally manipulated individuals. Cell Biol Int Rep. 1985 Dec;9(12):1069–1074. doi: 10.1016/s0309-1651(85)80004-7. [DOI] [PubMed] [Google Scholar]
- Tamone S. L., Chang E. S. Methyl farnesoate stimulates ecdysteroid secretion from crab Y-organs in vitro. Gen Comp Endocrinol. 1993 Mar;89(3):425–432. doi: 10.1006/gcen.1993.1050. [DOI] [PubMed] [Google Scholar]
- Wainwright G., Webster S. G., Wilkinson M. C., Chung J. S., Rees H. H. Structure and significance of mandibular organ-inhibiting hormone in the crab, Cancer pagurus. Involvement in multihormonal regulation of growth and reproduction. J Biol Chem. 1996 May 31;271(22):12749–12754. doi: 10.1074/jbc.271.22.12749. [DOI] [PubMed] [Google Scholar]