Purification and cDNA cloning of a cecropin-like peptide from the great wax moth, Galleria mellonella.
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.
Kim
Chong Han
CH
Department of Life Science, Hoseo University, Asan 337-850, Korea.
Lee
Joon Ha
JH
Kim
Iksoo
I
Seo
Sook Jae
SJ
Son
Seok Min
SM
Lee
Ki Young
KY
Lee
In Hee
IH
D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000302:isolation & purification* / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D005216:Fat Body / Q000737:chemistry; D006458:Hemolymph / Q000737:chemistry; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000033:anatomy & histology / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000890:Anti-Infective Agents; D019476:Insect Proteins; D010455:Peptides