Purification and cDNA cloning of a cecropin-like peptide from the great wax moth, Galleria mellonella

Mol Cells. 2004 Apr 30;17(2):262-6.

Abstract

A cecropin-like antimicrobial peptide, Gm cecropin, was purified from hemolymph of larvae of the wax moth, Galleria mellonella, immunized against E. coli, and its antibacterial activity was examined in a radial diffusion assay. The molecular mass of Gm cecropin was 4,160.69 Da by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis. The full-length cDNA of the Gm cecropin precursor was cloned by a combination of RT-PCR, based on the N-terminal sequence obtained by Edman degradation, and 5'-RACE-PCR. Analysis of the cDNA showed that cecropin is synthesized as a prepropeptide, with a putative 22-residue signal peptide, a 4-residue propeptide and a 39-residue mature peptide with a calculated mass of 4,344.18 Da the difference between the calculated and measured masses suggests that Gm cecropin is a 37-residue peptide generated by removal of the C-terminal residue and amidation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Anti-Infective Agents / isolation & purification*
  • Anti-Infective Agents / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • Fat Body / chemistry
  • Hemolymph / chemistry
  • Insect Proteins / genetics
  • Insect Proteins / metabolism*
  • Larva / anatomy & histology
  • Larva / metabolism
  • Molecular Sequence Data
  • Moths / genetics
  • Moths / metabolism*
  • Peptides / genetics
  • Peptides / isolation & purification*
  • Peptides / metabolism*
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • Anti-Infective Agents
  • Insect Proteins
  • Peptides