Computational proteomics analysis of Taphrina deformans for the identification of antifungal drug targets and validation with commercial fungicides

Abstract

Taphrina deformans is a plant-pathogenic fungus and a responsible agent for causing peach leaf curl disease. Taphrina deformans affects peach fruit production and contributes to global economic losses. Commercial fungicides may provide temporary relief; however, their overuse resulted in adverse environmental consequences as well as led to drug-resistant strains of T. deformans. Therefore, the discovery of novel drug targets for the future synthesis of antifungal drugs against Taphrina deformans is needed. Here we studied Taphrina deformans by computational proteomics approaches. The whole genome and proteome of T. deformans were subjected to subtractive proteomics, high-throughput virtual screening, and molecular dynamic simulations. We employed subtractive proteomics analysis of 4,659 proteins extracted from UniProtKB database; after filtering out homologous and non-essential proteins, we identified 189 essential ones, including nine that participated in the crucial metabolic pathways of the pathogen. These proteins were categorized as nuclear (n = 116), cytoplasmic (n = 37), and membrane (n = 36). Of those essential proteins, glutamate–cysteine ligase (GCL) emerged as one promising target due to its essential function for glutathione biosynthesis process which facilitates T. deformans survival and pathogenicity. To validate GCL as an antifungal target, virtual screening and molecular docking studies with various commercial fungicides were carried out to better characterize GCL as a drug target. The data showed strong binding affinities for polyoxin D, fluoxastrobin, trifloxystrobin, and azoxystrobin within the active site of GCL. Polyoxin D showed a strong affinity when the measured docking score was at -7.34 kcal/mol, while molecular dynamics simulations confirmed stable interactions (three hydrogen bonds, two hydrophobic bonds, and one salt bridge interaction), supporting our findings that GCL represents an excellent target for antifungal drug development efforts. The results showed that GCL, as an innovative target for future fungicide designs to combat T. deformans infections, provides an avenue toward creating more effective peach leaf curl disease treatments while mitigating environmental harm caused by its current use.