Proteomic Approaches to Identify Cold-Regulated Plasma Membrane Proteins
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Plasma membrane is the primary determinant of freezing tolerance in plants because of its central role in freeze–thaw cycle. Changes in plasma membrane protein composition have been one of the major research areas in plant cold acclimation. To obtain comprehensive profiles of the plasma membrane proteomes and their changes during the cold acclimation process, a plasma membrane purification method using a dextran–polyethylene glycol two polymer system and a mass spectrometry-based shotgun proteomics method using nano-LC-MS/MS for the plasma membrane proteins are described. The proteomic results obtained are further applied to label-free protein semiquantification.
Key wordsCold acclimation Plasma membrane Nano-LC-MS/MS Shotgun proteomics Label-free semiquantification In-solution digestion In-gel digestion
This work was supported in part by a Research Fellowship for Young Scientists (#247373 to D.T.) and Grants-in-Aid for Scientific Research (#22120003, #24370018, #17H03961 to M.U. and Y.K.) from JSPS, Japan.
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