Target validation and ligand development for a pathogenic fungal profilin, using a knock-down strain of pathogenic yeast Candida glabrata and structure-based ligand design.
The emergence of antifungal drug resistance is triggering vigorous searches for novel antifungal targets and lead compounds. In this study, we focused on fungal profilin, which is a small actin control protein sharing limited homology to human profilin. To validate its potentiality as a target, a profilin-conditional mutant of the pathogenic yeast Candida glabrata was constructed, using a regulatable Tet promoter, and its growth was monitored in vitro. Repression of profilin expression led to severe growth defect, demonstrating the potential of this protein as a novel antifungal target. Next, novel peptides binding to the active interface of profilin were designed by computer simulation. ELISA analysis showed that these peptides did bind to the wild-type profilin but bound less strongly to a profilin with amino acid substitutions at the active interface. Hence, we show here that profilin is a potential antifungal target and offer novel peptide ligands.
Medical Mycology Research Center (MMRC), Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8673, Japan. uenok5@restaff.chiba-u.jp
Keigo
K
Ueno
Yutaka
Y
Tamura
Hiroji
H
Chibana
D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology*; D041221:Candida glabrata / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D003198:Computer Simulation; D015198:Designer Drugs / Q000378:metabolism / Q000494:pharmacology*; D015195:Drug Design; D004797:Enzyme-Linked Immunosorbent Assay; D055785:Gene Knockdown Techniques; D006801:Humans; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology*; D051304:Profilins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011485:Protein Binding
D000935:Antifungal Agents; D015198:Designer Drugs; D010455:Peptides; D051304:Profilins