Isolation and identification of a cAMP generating peptide from the flesh fly, Neobellieria bullata (Diptera: Sarcophagidae)

Arch Insect Biochem Physiol. 1996;31(2):135-47. doi: 10.1002/(SICI)1520-6327(1996)31:2<135::AID-ARCH2>3.0.CO;2-Z.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies
  • Chromatography, High Pressure Liquid
  • Cyclic AMP / biosynthesis*
  • Diptera / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • Immunoblotting
  • Insect Proteins*
  • Male
  • Malpighian Tubules / metabolism
  • Manduca / metabolism*
  • Molecular Sequence Data
  • Molecular Weight
  • Proteins / chemistry
  • Proteins / isolation & purification*
  • Proteins / metabolism*
  • Sensitivity and Specificity

Substances

  • Antibodies
  • Insect Proteins
  • Proteins
  • cAMP generating peptide, flesh fly
  • Cyclic AMP