Orbitrap Astral-based proteome and phosphoproteome analysis identifies candidate proteins associated with the phosphatidate phosphatase MoPah1 in Magnaporthe oryzae

基于Orbitrap Astral的蛋白质组和磷酸化蛋白质组分析鉴定出稻瘟病菌中与磷脂酸磷酸酶MoPah1相关的候选蛋白

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Abstract

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.

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