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Updated: Feb 4, 2026
Author Spotlight: Advancing EVtrap for High-Throughput Proteomics in Disease Biomarker Discovery
Published on: October 27, 2023
Orbitrap Astral-based proteome and phosphoproteome analysis identifies candidate proteins associated with the
Juan Zhao1, Lili Yang2, Xinyi Shi3
1College of Chemistry and Life Sciences, Sichuan Provincial Key Laboratory for Development and Utilization of Characteristic Horticultural Biological Resources, Chengdu Normal University, Chengdu, 611130, China. zhaojuan201809@163.com.
Background:
Previous studies have shown that the phosphatidate phosphatase MoPah1 plays a critical role in Magnaporthe oryzae development and pathogenicity. The gene deletion mutant exhibits reduced virulence on rice leaves; however, the MoPah1 regulatory network remains unclear.
Result:
In this study, we applied a quantitative data-independent acquisition approach using the Orbitrap Astral instrument to analyze the gene deletion mutant. We identified 6,799 proteins and 15,682 phosphorylation sites in M. oryzae, with differentially abundant proteins primarily enriched in metabolic and autophagy pathways. Integrating phosphoproteomic and proteomic analyses, we found 72 overlapping proteins. Additionally, we employed glutathione S-transferase pull-down and yeast two-hybrid assays to screen and verify MoPah1-interacting proteins, identifying Pmk1.
Conclusions:
Our findings reveal candidate proteins in the MoPah1 regulatory network and the interaction between MoPah1 and Pmk1 in M. oryzae.
Insights
This study investigates the regulatory network of MoPah1, a key protein in Magnaporthe oryzae. Researchers identified interacting proteins, including Pmk1, revealing new insights into fungal pathogenicity.
Area of Science:
- Fungal biology
- Molecular mechanisms of pathogenicity
- Proteomics and phosphoproteomics
Background:
- Phosphatidate phosphatase MoPah1 is crucial for Magnaporthe oryzae development and pathogenicity.
- MoPah1 gene deletion mutants show reduced virulence on rice.
- The regulatory network of MoPah1 is not well understood.
Purpose of the Study:
- To elucidate the MoPah1 regulatory network in Magnaporthe oryzae.
- To identify proteins interacting with MoPah1.
- To understand the role of MoPah1 in fungal pathogenicity.
Main Methods:
- Quantitative data-independent acquisition proteomics using Orbitrap Astral.
- Proteomic and phosphoproteomic analyses of a MoPah1 gene deletion mutant.
- Glutathione S-transferase pull-down and yeast two-hybrid assays.
Main Results:
- Identified 6,799 proteins and 15,682 phosphorylation sites in M. oryzae.
- Differentially abundant proteins were enriched in metabolic and autophagy pathways.
- Identified 72 overlapping proteins between phosphoproteomic and proteomic analyses.
- Discovered Pmk1 as an interacting protein with MoPah1.
Conclusions:
- Candidate proteins in the MoPah1 regulatory network were revealed.
- The interaction between MoPah1 and Pmk1 in M. oryzae was confirmed.
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