RT Journal Article
T1 Phosphoproteome Analysis Using Two-Dimensional Electrophoresis Coupled with Chemical Dephosphorylation
A1 Rodríguez Vázquez, Raquel
A1 Mouzo Calzadilla, Daniel
A1 Zapata Babío, José Carlos
K1 Hydrogen fluoride-pyridine
K1 Meat phosphoproteome
K1 Phosphorylation
K1 Phosphorylation rate
K1 Post-translational protein modification
AB Protein phosphorylation is a reversible post-translational modification (PTM) with major regulatory roles in many cellular processes. However, the analysis of phosphoproteins remains the most challenging barrier in the prevailing proteome research. Recent technological advances in two-dimensional electrophoresis (2-DE) coupled to mass spectrometry (MS) have enabled the identification, characterization, and quantification of protein phosphorylation on a global scale. Most research on phosphoproteins with 2-DE has been conducted using phosphostains. Nevertheless, low-abundant and low-phosphorylated phosphoproteins are not necessarily detected using phosphostains and/or MS. In this study, we report a comparative analysis of 2-DE phosphoproteome profiles using Pro-Q Diamond phosphoprotein stain (Pro-Q DPS) and chemical dephosphorylation of proteins with HF-P from longissimus thoracis (LT) muscle samples of the Rubia Gallega cattle breed. We found statistically significant differences in the number of identified phosphoproteins between methods. More specifically, we found a three-fold increase in phosphoprotein detection with the HF-P method. Unlike Pro-Q DPS, phosphoprotein spots with low volume and phosphorylation rate were identified by HF-P technique. This is the first approach to assess meat phosphoproteome maps using HF-P at a global scale. The results open a new window for 2-DE gel-based phosphoproteome analysis.
PB MDPI
YR 2022
FD 2022-10-07
LK https://hdl.handle.net/10347/46128
UL https://hdl.handle.net/10347/46128
LA eng
NO Rodríguez-Vázquez, R., Mouzo, D., & Zapata, C. (2022). Phosphoproteome Analysis Using Two-Dimensional Electrophoresis Coupled with Chemical Dephosphorylation. Foods, 11(19), 3119. https://doi.org/10.3390/foods11193119
NO This work was supported by the Instituto Nacional de Investigación y Tecnología Agraria (INIA, RTA 2014-00034-C04), Spain; and by a predoctoral fellowship of the Xunta de Galicia (Spain) and the European Union (ESF) to R. Rodríguez-Vázquez.
DS Minerva
RD 23 may 2026