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000279386 1001_ $$0P:(DE-Juel1)131953$$aBratanov, Dmitry$$b0$$ufzj
000279386 245__ $$aAn Approach to Heterologous Expression of Membrane Proteins. The Case of Bacteriorhodopsin
000279386 260__ $$aLawrence, Kan.$$bPLoS$$c2015
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000279386 520__ $$aHeterologous overexpression of functional membrane proteins is a major bottleneck of structural biology. Bacteriorhodopsin from Halobium salinarum (bR) is a striking example of the difficulties in membrane protein overexpression. We suggest a general approach with a finite number of steps which allows one to localize the underlying problem of poor expression of a membrane protein using bR as an example. Our approach is based on constructing chimeric proteins comprising parts of a protein of interest and complementary parts of a homologous protein demonstrating advantageous expression. This complementary protein approach allowed us to increase bR expression by two orders of magnitude through the introduction of two silent mutations into bR coding DNA. For the first time the high quality crystals of bR expressed in E. Coli were obtained using the produced protein. The crystals obtained with in meso nanovolume crystallization diffracted to 1.67 Å.
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000279386 7001_ $$0P:(DE-Juel1)131949$$aBalandin, Taras$$b1$$ufzj
000279386 7001_ $$0P:(DE-HGF)0$$aRound, Ekaterina$$b2
000279386 7001_ $$0P:(DE-Juel1)143908$$aShevchenko, Vitaly$$b3$$ufzj
000279386 7001_ $$0P:(DE-Juel1)165798$$aGushchin, Ivan$$b4$$ufzj
000279386 7001_ $$0P:(DE-HGF)0$$aPolovinkin, Vitaly$$b5
000279386 7001_ $$0P:(DE-Juel1)144613$$aBorshchevskiy, Valentin$$b6
000279386 7001_ $$0P:(DE-Juel1)131964$$aGordeliy, Valentin$$b7$$eCorresponding author$$ufzj
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