Isolation and characterization of Melanoplus sanguinipes adipokinetic hormone: a new member of the AKH/RPCH family.
A neuropeptide hormone isolated from corpora cardiaca of Melanoplus sanguinipes was purified by HPLC. The HPLC fractions were examined for adipokinetic activity with an in vivo bioassay. A single large UV absorbent peak was active in the mobilization of lipid while the other HPLC fractions showed no detectable activity. This large peak had a retention time and amino acid composition identical to synthetic Lom-AKH-I which was analyzed in a parallel manner. The primary sequence structure, pGlu-Leu-Asn-Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2, was determined by automated gas-phase Edman degradation. The peptide was deblocked prior to sequencing using pyroglutamate aminopeptidase and the sequence was confirmed with mass spectrometry. The C-terminus of the peptide was determined to be blocked, as indicated by the lack of digestion with carboxypeptidase A. The knowledge of the primary sequence of Mes-AKH allows the use of a commercially available synthetic peptide and its antibodies for use in future research with Melanoplus sanguinipes.
Taub-Montemayor
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Department of Zoology, University of Texas at Austin 78712-1064, USA. ttaub@mail.utexas.edu
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D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006110:Grasshoppers; D007301:Insect Hormones / Q000737:chemistry* / Q000302:isolation & purification*; D009479:Neuropeptides / Q000737:chemistry / Q000302:isolation & purification; D009842:Oligopeptides / Q000737:chemistry* / Q000302:isolation & purification*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007301:Insect Hormones; D009479:Neuropeptides; D009842:Oligopeptides; C044833:adipokinetic hormone; D011761:Pyrrolidonecarboxylic Acid