Purification and chemical characterization of peptide G1, an invertebrate neuropeptide that stimulates cyclic GMP metabolism.
Bioassay analysis of extracts of the major neurosecretory structures of the American lobster have revealed several different agents with stimulatory effects on the cyclic GMP metabolism of various lobster tissues. The most potent of these is a peptide extracted from the sinus gland, a neurohemal organ found in the animal's eyestalk. This molecule, called peptide G1 (for its effects on cyclic GMP metabolism), can increase the cyclic GMP content of every lobster tissue tested, sometimes by as much as 200-fold. In this article, we describe the purification and some of the chemical properties of peptide G1. Purification was accomplished by sequential anion exchange and reverse-phase HPLC. The purified peptide is a large, extremely hydrophobic molecule. Its apparent molecular mass on a reducing sodium dodecyl sulfate-containing gel is 6.4 kDa, and its calculated molecular mass (based on an amino acid analysis of the purified material) is 8.2 kDa. Amino acid analysis reveals a high proportion of leucine and valine residues. The amino terminus of the molecule is not susceptible to Edman degradation, but sequencing studies were successfully carried out on tryptic fragments. Based on the estimated size of the molecule, these studies provide approximately 60% of the total sequence. No homologies with any previously sequenced peptide were observed, but biochemical similarities to as yet unsequenced peptides found in extracts of sinus glands from other crustaceans (hyperglycemic hormone and moult-inhibiting hormone) are described.
Pavloff
M S
MS
Department of Physiology, University of North Carolina, Chapel Hill 27599-7545.
Goy
M F
MF
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D007447:Invertebrate Hormones / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D008121:Nephropidae / Q000032:analysis*; D009479:Neuropeptides / Q000302:isolation & purification*; D009490:Neurosecretory Systems / Q000032:analysis; D010446:Peptide Fragments; D012689:Sequence Homology, Nucleic Acid; D014357:Trypsin
D000596:Amino Acids; D007447:Invertebrate Hormones; D009479:Neuropeptides; D010446:Peptide Fragments; D014357:Trypsin