TY - JOUR
T1 - Expression and purification of the SNX1/SNX6 complex
AU - Yong, Xin
AU - Hu, Wenfeng
AU - Zhou, Xue
AU - Wang, Jing
AU - Burstein, Ezra
AU - Jia, Da
N1 - Funding Information:
We thank members of our laboratory for helpful discussions. Research is supported by Natural Science Foundation of China (NSFC) grants ( #31671477 to D.J.). D.J. is a Chinese ‘‘One Thousand Talents’’ program scholar.
Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2018/11
Y1 - 2018/11
N2 - 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.
AB - 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.
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U2 - 10.1016/j.pep.2018.06.010
DO - 10.1016/j.pep.2018.06.010
M3 - Article
C2 - 29908913
AN - SCOPUS:85048854444
SN - 1046-5928
VL - 151
SP - 93
EP - 98
JO - Protein Expression and Purification
JF - Protein Expression and Purification
ER -