Proteomics of Endosomal Compartments from Plants Case Study: Isolation of Trans-Golgi Network Vesicles
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A detailed understanding of endomembrane processes and their biological roles is vital for a complete picture of plant growth and development; however their highly dynamic nature has complicated comprehensive and rigorous studies so far. Recent pioneering efforts have demonstrated that isolation of vesicles in their native state, paired with a quantitative identification of their cargo, offers a viable and practicable approach for the dissection of endomembrane trafficking pathways. The protocol presented in this chapter describes in detail the isolation of the SYP61 trans-Golgi network vesicles from Arabidopsis. With minor alterations, in a few key parameters, it can be adopted to yield a universal procedure for the broad spectrum of plant vesicles.
Key wordsVesicle isolation Immunoisolation Trans-Golgi network (TGN) SYP61 Endomembrane trafficking Proteomics
This work was supported by the UCD Hellman Fellowship and the NSF-IOS 1258135 to GD.
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