Journal Article

FZJ-2015-07398

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png An Approach to Heterologous Expression of Membrane Proteins. The Case of Bacteriorhodopsin

Bratanov, D.FZJ* ; Balandin, T.FZJ* ; Round, E. ; Shevchenko, V.FZJ* ; Gushchin, I.FZJ* ; Polovinkin, V. ; Borshchevskiy, V. ; Gordeliy, V. (Corresponding author)FZJ*

2015
PLoS Lawrence, Kan.

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Please use a persistent id in citations: http://hdl.handle.net/2128/9572  doi:10.1371/journal.pone.0128390

Abstract: Heterologous overexpression of functional membrane proteins is a major bottleneck of structural biology. Bacteriorhodopsin from Halobium salinarum (bR) is a striking example of the difficulties in membrane protein overexpression. We suggest a general approach with a finite number of steps which allows one to localize the underlying problem of poor expression of a membrane protein using bR as an example. Our approach is based on constructing chimeric proteins comprising parts of a protein of interest and complementary parts of a homologous protein demonstrating advantageous expression. This complementary protein approach allowed us to increase bR expression by two orders of magnitude through the introduction of two silent mutations into bR coding DNA. For the first time the high quality crystals of bR expressed in E. Coli were obtained using the produced protein. The crystals obtained with in meso nanovolume crystallization diffracted to 1.67 Å.

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Contributing Institute(s):

1. Strukturbiochemie (ICS-6)

Research Program(s):

1. 551 - Functional Macromolecules and Complexes (POF3-551) (POF3-551)

Appears in the scientific report 2015

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Database coverage:
; ; ; ; BIOSIS Previews ; IF < 5 ; JCR ; NCBI Molecular Biology Database ; SCOPUS ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection ; Zoological Record

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