The synthesis of recombinant membrane proteins in yeast for structural studies

Routledge, Sarah, Mikaliunaite, Lina, Patel, Anjana, Clare, Michelle, Cartwright, Stephanie P., Bawa, Zharain, Wilks, Martin D.B., Low, Floren, Hardy, David, Rothnie, Alice J. and Bill, Roslyn M. (2016). The synthesis of recombinant membrane proteins in yeast for structural studies. Methods, 95 , pp. 26-37.

Abstract

Historically, recombinant membrane protein production has been a major challenge meaning that many fewer membrane protein structures have been published than those of soluble proteins. However, there has been a recent, almost exponential increase in the number of membrane protein structures being deposited in the Protein Data Bank. This suggests that empirical methods are now available that can ensure the required protein supply for these difficult targets. This review focuses on methods that are available for protein production in yeast, which is an important source of recombinant eukaryotic membrane proteins. We provide an overview of approaches to optimize the expression plasmid, host cell and culture conditions, as well as the extraction and purification of functional protein for crystallization trials in preparation for structural studies.

Publication DOI: https://doi.org/10.1016/j.ymeth.2015.09.027
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Additional Information: Funding: BBSRC (grants BB/I019960/1, BB/K013319/1, BB/L502194/1 and BB/L015846/1; and the Innovative Medicines Joint Undertaking (grant 115583 to the ND4BB ENABLE Consortium)
Uncontrolled Keywords: membrane proteins,yeast,recombinant antigen,expression plasmid vector,promoter,SMALP,cholesterol,Molecular Biology,Biochemistry, Genetics and Molecular Biology(all)
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2016-02-15
Authors: Routledge, Sarah
Mikaliunaite, Lina
Patel, Anjana
Clare, Michelle
Cartwright, Stephanie P.
Bawa, Zharain
Wilks, Martin D.B.
Low, Floren
Hardy, David
Rothnie, Alice J. ( 0000-0002-4259-7015)
Bill, Roslyn M. ( 0000-0003-1331-0852)

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