4.5 Article

Expression of recombinant eyestalk crustacean hyperglycemic hormone from the tropical land crab, Gecarcinus lateralis, that inhibits Y-organ ecdysteroidogenesis in vitro

期刊

MOLECULAR BIOLOGY REPORTS
卷 36, 期 6, 页码 1231-1237

出版社

SPRINGER
DOI: 10.1007/s11033-008-9302-8

关键词

Crustacea; Arthropoda; Hormone; Neuropeptide; Ecdysteroid; Y-organ; Gene expression; Crustacean hyperglycemic hormone; Molting; DNA sequence; Amino acid sequence; cDNA; Cloning; Yeast expression system; Pichia pastoris

资金

  1. National Science Foundation [IBN-0342982]

向作者/读者索取更多资源

Crustacean hyperglycemic hormone (CHH) is a pleiotropic neuropeptide that regulates carbohydrate and lipid metabolism, molting, reproduction, and osmoregulation in decapod crustaceans. CHH elevates glucose levels in the hemolymph by stimulating glycogenolysis in target tissues. It also inhibits ecdysteroidogenesis in the molting gland, or Y-organ (YO), possibly as a response to environmental stress. CHH acts via binding to a membrane receptor guanylyl cyclase, which is expressed in most tissues, including the YO. Large amounts of biologically active neuropeptide are required to investigate the mechanism of CHH signaling in the YO. Consequently, the eyestalk ganglia CHH (EG-CHH) isoform was cloned into a yeast (Pichia pastoris) expression vector to express recombinant mature peptide (rEG-CHH) with or without a C-terminal c-Myc/polyhistidine tag. Yeast cultures with untagged or tagged rEG-CHH inhibited ecdysteroidogenesis in YOs from European green crab (Carcinus maenas) 36% (P < 0.002) and 51% (P < 0.006), respectively. Purified tagged EG-CHH inhibited YO ecdysteroidogenesis 32% (P < 0.002), but lacked hyperglycemic activity in vivo. This is the first report of recombinant EG-CHH inhibiting YO ecdysteroidogenesis. The data suggest that the tagged recombinant peptide can be used to elucidate the CHH signaling pathway in the crustacean molting gland.

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