The active site of drosomycin, a small insect antifungal protein, delineated by comparison with the modeled structure of Rs-AFP2, a plant antifungal protein.
Drosomycin is the first strictly antifungal protein isolated from an insect (Drosophila melanogaster). The solution structure of this 44-residue protein has been reported previously. It involves a three-stranded beta-sheet and an alpha-helix, the protein global fold being maintained by four disulfide bridges. Rs-AFP2 is a plant antifungal protein exhibiting 41% sequence similarity with drosomycin. Mutational analysis of Rs-AFP2 showed the importance of some residues in the antifungal activity of the protein against the fungus target. In order to determine the structural features responsible for antifungal activity in both drosomycin and Rs-AFP2, we modeled the three-dimensional structure of Rs-AFP2, and of other antifungal proteins, using the solution structure of drosomycin as a template. Structure analysis of drosomycin and Rs-AFP2, and comparisons with the other modeled antifungal structures, revealed that the two proteins shared a hydrophobic cluster located at the protein surface in which a lysine residue is embedded. Based on these close structural similarities and the experimental data available for Rs-AFP2 mutants, an antifungal active site of the insect protein is proposed.
Landon
C
C
Centre de Biophysique Moléculaire, CNRS-UPR 4301, Orléans University, France.
Pajon
A
A
Vovelle
F
F
Sodano
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000935:Antifungal Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides; D001665:Binding Sites; D023082:Defensins; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D008239:Lysine / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D010944:Plants / Q000737:chemistry; D017433:Protein Structure, Secondary; D016415:Sequence Alignment
D000935:Antifungal Agents; D023181:Antimicrobial Cationic Peptides; D023082:Defensins; D029721:Drosophila Proteins; D019476:Insect Proteins; D010940:Plant Proteins; C091227:DRS protein, Drosophila; D008239:Lysine