Structural and biological characterization of chromofungin, the antifungal chromogranin A-(47-66)-derived peptide.
Vasostatin-I, the natural fragment of chromogranin A-(1-76), is a neuropeptide able to kill a large variety of fungi and yeast cells in the micromolar range. We have examined the antifungal properties of synthetic vasostatin-I-related peptides. The most active shortest peptide, named chromofungin, corresponds to the sequence Arg(47)-Leu(66). Extensive (1)H NMR analysis revealed that it adopts a helical structure. The biophysical mechanism implicated in the interaction of chromofungin with fungi and yeast cells was studied, showing the penetration of this peptide with different lipid monolayers. In order to examine thoroughly the antifungal activity of chromofungin, confocal laser microscopy was used to demonstrate the ability of the rhodamine-labeled peptide to interact with the fungal cell wall, to cross the plasma membrane, and to accumulate in Aspergillus fumigatus, Alternaria brassicola, and Candida albicans. Our present data reveal that chromofungin inhibits calcineurin activity, extending a previous observation that the N-terminal region of chromogranin A interacts with calmodulin in the presence of calcium. Therefore, the destabilization of fungal wall and plasma membrane, together with the possible intracellular inhibition of calmodulin-dependent enzymes, is likely to represent the mechanism by which vasostatin-I and chromofungin exert antifungal activity.
Lugardon
K
K
INSERM Unité 338, IFR37 "Biologie de la Communication Cellulaire," 5 rue Blaise Pascal 67084 Strasbourg Cedex, France.
Chasserot-Golaz
S
S
Kieffer
A E
AE
Maget-Dana
R
R
Nullans
G
G
Kieffer
B
B
Aunis
D
D
Metz-Boutigue
M H
MH
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry* / Q000494:pharmacology*; D053379:Chromogranin A; D002864:Chromogranins / Q000737:chemistry* / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D018613:Microscopy, Confocal; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
D000935:Antifungal Agents; D053379:Chromogranin A; D002864:Chromogranins; D010446:Peptide Fragments; C437333:chromofungin