Label-Free Proteomic Analysis of Molecular Effects of 2-Methoxy-1,4-naphthoquinone on Penicillium italicum

Abstract

Penicillium italicum is the principal pathogen causing blue mold of citrus. Searching for novel antifungal agents is an important aspect of the post-harvest citrus industry because of the lack of higher effective and low toxic antifungal agents. Herein, the effects of 2-methoxy-1,4-naphthoquinone (MNQ) on P. italicum and its mechanism were carried out by a series of methods. MNQ had a significant anti-P. italicum effect with an MIC value of 5.0 µg/mL. The label-free protein profiling under different MNQ conditions identified a total of 3037 proteins in the control group and the treatment group. Among them, there were 129 differentially expressed proteins (DEPs，up-regulated > 2.0-fold or down-regulated < 0.5-fold, p < 0.05), 19 up-regulated proteins, 26 down-regulated proteins, and 67 proteins that were specific for the treatment group and another 17 proteins that were specific for the control group. Of these, 83 proteins were sub-categorized into 23 hierarchically-structured GO classifications. Most of the identified DEPs were involved in molecular function (47%), meanwhile 27% DEPs were involved in the cellular component and 26% DEPs were involved in the biological process. Twenty-eight proteins identified for differential metabolic pathways by KEGG were sub-categorized into 60 classifications. Functional characterization by GO and KEGG enrichment results suggests that the DEPs are mainly related to energy generation (mitochondrial carrier protein, glycoside hydrolase, acyl-CoA dehydrogenase, and ribulose-phosphate 3-epimerase), NADPH supply (enolase, pyruvate carboxylase), oxidative stress (catalase, glutathione synthetase), and pentose phosphate pathway (ribulose-phosphate 3-epimerase and xylulose 5-phosphate). Three of the down-regulated proteins selected randomly the nitro-reductase family protein, mono-oxygenase, and cytochrome P450 were verified using parallel reaction monitoring. These findings illustrated that MNQ may inhibit P. italicum by disrupting the metabolic processes, especially in energy metabolism and stimulus response that are both critical for the growth of the fungus. In conclusion, based on the molecular mechanisms, MNQ can be developed as a potential anti-fungi agent against P. italicum.

Identifying Potential Mitochondrial Proteome Signatures Associated with the Pathogenesis of Pulmonary Arterial Hypertension in the Rat Model

“…We used Orbitrap Fusion Lumos (Thermo Scientific) to perform label-free quantitative analysis, coupled with an Easy nLC/Ultimate 3000 System (Thermo Scientific). Analysis of peptide information was adopted by liquid chromatography- (LC-) electrospray ionization (ESI) tandem MS (MS/MS) analysis as previously described [ 23 , 24 ].…”

Physiological and Proteomic Analysis of Penicillium digitatum in Response to X33 Antifungal Extract Treatment

“…Accumulation of ROS is able to affects the normal cellular and membrane functions through oxidizing lipids, nucleic acids, proteins, and carbohydrates (Guo et al, 2019). Phosphatidylcholine and ergosterol are two of the major components of the P. digitatum cell membrane.…”

Label‐free proteomic dissection on dptP‐deletion mutant uncovers dptP involvement in strain growth and daptomycin tolerance of Streptomyces roseosporus

“…Proteomics technology provides a promising exploitation in natural product discovery (Acharya et al ., 2019), post‐translational modification (Sun et al ., 2020), elucidation of biosynthetic pathway (Bordoloi et al ., 2016), even illustration of molecular mechanism response to a certain treatment (Guo et al ., 2019). Specially, LFQ‐based proteomic approach is simple and time saving for sample preparation and result analyses of MS identification and quantification.…”