TY  - JOUR
AU  - Bratanov, Dmitry
AU  - Balandin, Taras
AU  - Round, Ekaterina
AU  - Shevchenko, Vitaly
AU  - Gushchin, Ivan
AU  - Polovinkin, Vitaly
AU  - Borshchevskiy, Valentin
AU  - Gordeliy, Valentin
TI  - An Approach to Heterologous Expression of Membrane Proteins. The Case of Bacteriorhodopsin
JO  - PLoS one
VL  - 10
IS  - 6
SN  - 1932-6203
CY  - Lawrence, Kan.
PB  - PLoS
M1  - FZJ-2015-07398
SP  - e0128390 -
PY  - 2015
AB  - 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.
LB  - PUB:(DE-HGF)16
UR  - <Go to ISI:>//WOS:000355652200066
C6  - pmid:26046789
DO  - DOI:10.1371/journal.pone.0128390
UR  - https://juser.fz-juelich.de/record/279386
ER  -