Expression and purification of the SNX1/SNX6 complex

Xin Yong, Wenfeng Hu, Xue Zhou, Jing Wang, Ezra Burstein, Da Jia

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Abstract

The sorting nexin (SNX) family proteins play an essential role in vesicular transport, cell signaling, and membrane remodeling. The SNX members SNX1/2 and SNX5/6 form dimers, and mediate endosome-to-trans Golgi network (TGN) transport through coordinating cargo selection and membrane remodeling. It is well-known how a SNX-BAR protein forms a homodimer; however, it is less clear how a heterodimer is formed. Here a detailed expression and purification protocol of the SNX1/SNX6 complex, from both worm and human, is described. Keys to the successful protein production include co-expression of both genes, and inclusion of glycerol in the protein buffer. Solution studies suggest that SNX1 and SNX6 form a 1:1 heterodimer. The production of a large amount, high quality of the SNX1/SNX6 complex provides a basis for future biochemical and structural studies of the complex, and in vitro reconstitution of SNX1/SNX6-mediated transport.

Original languageEnglish (US)
Pages (from-to)93-98
Number of pages6
JournalProtein Expression and Purification
Volume151
DOIs
Publication statusPublished - Nov 1 2018

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ASJC Scopus subject areas

  • Biotechnology

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