Volume 22, No 2, Feb 2012

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 22 Issue 2, February 2012: 413-424

ORIGINAL ARTICLES

Isolation and proteomic analysis of the SYP61 compartment reveal its role in exocytic trafficking in Arabidopsis

Georgia Drakakaki^1,6, Wilhelmina van de Ven¹, Songqin Pan¹, Yansong Miao^2,7, Junqi Wang², Nana K Keinath³, Brent Weatherly^4,5, Liwen Jiang², Karin Schumacher

¹Department of Botany and Plant Sciences, Center for Plant Cell Biology, Institute for Integrative Genome Biology, 4119C Genomics Building, University of California Riverside, CA 92521, USA
²School of Life Sciences, Center for Cell and Developmental Biology, Chinese University of Hong Kong, New Territories, Hong Kong, China
³Heidelberg Institute for Plant Science, Im Neuenheimer Feld 230, Heidelberg 69120, Germany
⁴Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA
⁵NuSep Inc., Bogart, GA 30622, USA
⁶Current address: Department of Plant Sciences, University of California, Davis, CA 95616, USA
⁷Current address: Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA Correspondence: Natasha Raikhel,(nraikhel@ucr.edu)

The endomembrane system is a complex and dynamic intracellular trafficking network. It is very challenging to track individual vesicles and their cargos in real time; however, affinity purification allows vesicles to be isolated in their natural state so that their constituent proteins can be identified. Pioneering this approach in plants, we isolated the SYP61 trans-Golgi network compartment and carried out a comprehensive proteomic analysis of its contents with only minimal interference from other organelles. The proteome of SYP61 revealed the association of proteins of unknown function that have previously not been ascribed to this compartment. We identified a complete SYP61 SNARE complex, including regulatory proteins and validated the proteome data by showing that several of these proteins associated with SYP61 in planta. We further identified the SYP121-complex and cellulose synthases, suggesting that SYP61 plays a role in the exocytic trafficking and the transport of cell wall components to the plasma membrane. The presence of proteins of unknown function in the SYP61 proteome including ECHIDNA offers the opportunity to identify novel trafficking components and cargos. The affinity purification of plant vesicles in their natural state provides a basis for further analysis and dissection of complex endomembrane networks. The approach is widely applicable and can afford the study of several vesicle populations in plants, which can be compared with the SYP61 vesicle proteome.

Cell Research (2012) 22:413-424. doi:10.1038/cr.2011.129; published online 9 August 2011

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