A novel proteomics workflow for simultaneous analysis of protein phosphorylation and S-nitrosylationOA
Protein post-translational modifications such as phosphorylation and S-nitrosylation regulate protein functions and cellular programs in eukaryotes.Moreover,extensive evidence suggests crosstalk between these modifications.However,we lack a comprehensive method for the simultaneous detection and analysis of multiple post-translational modifications.Here,we present an optimized workflow that identifies phosphorylation and S-nitrosylation sites using a novel phosphate affinity tag switch technique.Validation with model proteins and complex biological samples confirmed the high sensitivity,coverage,and reproducibility of this method.Applying this method to Arabidopsis thaliana seedlings revealed 12,552 phosphorylation sites and 6,108 S-nitrosylation sites,representing the largest single-study dataset of S-nitrosylation sites to date.This approach enhances our understanding of post-translational modification dynamics in plant signaling,stress responses,and metabolism.
Wenyang Zhang;Yanjiao Wang;Wenyan Li;Shaowen Wu;Yuanyuan Chen;Mingyang Ye;Wenjie Huang;Alisdair R.Fernie;Shijuan Yan
State Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources,Guangdong Laboratory for Lingnan Modern Agriculture,College of Life Sciences,South China Agricultural University,Guangzhou 510642,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources,Guangdong Laboratory for Lingnan Modern Agriculture,College of Life Sciences,South China Agricultural University,Guangzhou 510642,ChinaState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,ChinaMax Planck Institute of Molecular Plant Physiology,Am Muhlenberg 1,14476 Potsdam-Golm,GermanyState Key Laboratory of Swine and Poultry Breeding Industry,Guangdong Key Laboratory for Crop Germplasm Resources Preservation and Utilization,Agro-Biological Gene Research Center,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China
生物科学
ProteomicsS-nitrosylationPhosphorylationPTM crosstalk
《aBIOTECH》 2025 (3)
P.452-465,14
support of the Science and Technology Program of Guangdong Province(2023A0505090005,2021TQ06N115)the Natural Science Foundation of Guangdong Province(2022A1515110962)the Special Fund for Scientific Innovation Strategy-Construction of High-Level Academy of Agriculture Science(R2021YJ-YB3010,R2023PY-JG025,2023QZ-NK04,GDNKY-ZQQZ-K5)Modern Seed Industry Innovation Capability Enhancement Project of Guangdong Academy of Agricultural Sciences.
评论