The molt-inhibiting hormone in the American crayfish Procambarus clarkii: its chemical synthesis and biological activity.
The molt-inhibiting hormone of the American crayfish Procambarus clarkii (Prc-MIH), a 75-residue polypeptide containing three disulfide bridges, was synthesized by chemical ligation of two peptides, i.e., synthetic Prc-MIH(1-39) and Prc-MIH(40-75)-NH(2), and by subsequent folding to form the native disulfide-containing peptide molecule. The synthetic peptide was comparable to the natural Prc-MIH in inhibiting ecdysteroid secretion by in vitro bioassay and shared features with the natural Prc-MIH in some biochemical analyses. These results indicate that the chemical ligation method can be used for the synthesis of Prc-MIH. Furthermore, it was demonstrated that synthetic Prc-MIH has hyperglycemic activity, although the activity was weaker than that of the authentic crustacean hyperglycemic hormone in the American crayfish. To examine the structural requirement of the Prc-MIH for eliciting biological activity, an antibody raised against the C-terminal region (residues 55-75) and two synthetic peptides, i.e., a core region (residues 1-54) containing three disulfide bridges and the C-terminal region, were utilized. It is suggested that Prc-MIH exerts its activities through coordination between the core region and the C-terminal region.
Sonobe
H
H
Department of Biology, Konan University, Kobe 658-8501, Japan. sonobe@konan-u.ac.jp
Nishimura
T
T
Sonobe
M
M
Nakatsuji
T
T
Yanagihara
R
R
Kawakami
T
T
Aimoto
S
S
D000818:Animals; D003400:Astacoidea / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000737:chemistry; D006868:Hydrolysis; D007447:Invertebrate Hormones / Q000138:chemical synthesis* / Q000502:physiology*; D009479:Neuropeptides / Q000138:chemical synthesis* / Q000502:physiology*; D010455:Peptides / Q000138:chemical synthesis / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004220:Disulfides; D007447:Invertebrate Hormones; D009479:Neuropeptides; D010455:Peptides; C012445:molt-inhibiting hormone