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Isolation and identification of a cAMP generating peptide from the flesh fly, Neobellieria bullata (Diptera: Sarcophagidae)
Author(s) -
Spittaels Kurt,
Devreese Bart,
Schoofs Liliane,
Neven Hedwig,
Janssen Ine,
Grauwels Luc,
Van Beeumen Jozef,
De Loof Arnold
Publication year - 1996
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/(sici)1520-6327(1996)31:2<135::aid-arch2>3.0.co;2-z
Subject(s) - flesh fly , biology , flesh , isolation (microbiology) , zoology , insect , botany , microbiology and biotechnology , horticulture , larva
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 μg Neb‐cGP and that 10 μg represents the lowest immunostainable amount on a Western blot. © 1996 Wiley‐Liss, Inc.