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@ARTICLE{Bratanov:279386,
author = {Bratanov, Dmitry and Balandin, Taras and Round, Ekaterina
and Shevchenko, Vitaly and Gushchin, Ivan and Polovinkin,
Vitaly and Borshchevskiy, Valentin and Gordeliy, Valentin},
title = {{A}n {A}pproach to {H}eterologous {E}xpression of
{M}embrane {P}roteins. {T}he {C}ase of {B}acteriorhodopsin},
journal = {PLoS one},
volume = {10},
number = {6},
issn = {1932-6203},
address = {Lawrence, Kan.},
publisher = {PLoS},
reportid = {FZJ-2015-07398},
pages = {e0128390 -},
year = {2015},
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.},
cin = {ICS-6},
ddc = {500},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551)},
pid = {G:(DE-HGF)POF3-551},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000355652200066},
pubmed = {pmid:26046789},
doi = {10.1371/journal.pone.0128390},
url = {https://juser.fz-juelich.de/record/279386},
}