title: Isolation and identification of a cAMP generating peptide from the flesh fly, Neobellieria bullata (Diptera: Sarcophagidae).
abstract: The Manduca sexta Malpighian tubule assay system, developed to monitor adenylate cyclase activity, was used in combination with HPLC to isolate a novel cAMP generating peptide from 350,000 whole flesh flies, Neobellieria bullata. Mass spectrometry revealed a molecular mass of 5,047 daltons, and Edman degradation the following sequence: AGAEAEKLSGLSKYFNGTTMAGRANVAKATYAVIGLIIAYNVMKPKKK. This 48-mer peptide, called Neb-cGP, does not belong to the corticotropin releasing factor family of insect diuretic peptides. Electrophoresis and subsequent immunoblotting of peptides immunoprecipitated from a homogenate of entire flies showed that one fly contained approximately 0.003 to 0.03 micrograms Neb-cGP and that 10 micrograms represents the lowest immunostainable amount on a Western blot.
authors.affiliation: Zoologisch Institut, Katholieke Universiteit Leuven, Belgium.
authors.forename: K
authors.initials: K
authors.lastname: Spittaels
authors.forename: B
authors.initials: B
authors.lastname: Devreese
authors.forename: L
authors.initials: L
authors.lastname: Schoofs
authors.forename: H
authors.initials: H
authors.lastname: Neven
authors.forename: I
authors.initials: I
authors.lastname: Janssen
authors.forename: L
authors.initials: L
authors.lastname: Grauwels
authors.forename: J
authors.initials: J
authors.lastname: Van Beeumen
authors.forename: A
authors.initials: A
authors.lastname: De Loof
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000096:biosynthesis*; D004175:Diptera / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015151:Immunoblotting; D019476:Insect Proteins; D008297:Male; D008317:Malpighian Tubules / Q000378:metabolism; D018402:Manduca / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies; D019476:Insect Proteins; D011506:Proteins; C098180:cAMP generating peptide, flesh fly; D000242:Cyclic AMP