Molecular and cellular endocrinology vol:117 issue:2 pages:157-165
A methanolic extract of 350000 adult grey fleshflies Neobellieria bullata, was prepared and screened for myotropic activity. After fractionation on the first column, all fractions were screened in two heterologous (Locusta oviduct and Leucophaea hindgut) and one homologous (Neobellieria hindgut) myotropic bioassay. We here report the purification of one fraction, which stimulates the contractions of the Locusta oviduct. Electrospray Mass Spectrometry of the peptide revealed a molecular mass of 1395.82. The primary structure has been determined as AYRKPPFNGSLF-amide. This novel peptide was designated Neb-LFamide. This sequence is different from the other known myotropic peptides in insects. The threshold concentration of the synthetic peptide is 1 x 10(-7) M on the Locusta oviduct. On the hindgut of Neobellieria in or Leucophaea, the synthetic peptide is not active. By use of a polyclonal antiserum raised against the synthetic peptide, immunoreactivity was localized in median neurosecretory cells in the pars intercerebralis of the fly brain, indicating that Neb-LFamide is a neuropeptide.