TY - JOUR
T1 - Expression of microbial rhodopsins in Escherichia coli and their extraction and purification using styrene-maleic acid copolymers
AU - Kojima, Keiichi
AU - Sudo, Yuki
N1 - Funding Information:
We thank Mr. Tetsuya Ueta, Dr. Tomoya Hino, Dr. Shingo Nagano and Dr. Mikihiro Shibata for sample preparation and invaluable discussion. This work was financially supported by JSPS KAKENHI grant numbers JP21K15054 to K.K. and JP20K21482 , JP21H02446, and JP21H00404 to Y.S. This research was partially supported by CREST -JST ( JPMJCR1656 ) to Y.S.
Funding Information:
We thank Mr. Tetsuya Ueta, Dr. Tomoya Hino, Dr. Shingo Nagano and Dr. Mikihiro Shibata for sample preparation and invaluable discussion. This work was financially supported by JSPS KAKENHI grant numbers JP21K15054 to K.K. and JP20K21482, JP21H02446, and JP21H00404 to Y.S. This research was partially supported by CREST-JST (JPMJCR1656) to Y.S. K.K. and Y.S. performed the research and wrote the manuscript. The authors declare no competing interests.
Publisher Copyright:
© 2021
PY - 2022/3/18
Y1 - 2022/3/18
N2 - Microbial rhodopsins are photoreceptive membrane proteins showing various light-dependent biological activities. Styrene-maleic acid (SMA) copolymers spontaneously form nanoscale lipid particles containing membrane proteins and associated lipids without detergent, and can be used to characterize membrane molecules. Here, we provide a protocol to functionally express a thermally stable rhodopsin, Rubrobacter xylanophilus rhodopsin, and an unstable rhodopsin, Halobacterium salinarum sensory rhodopsin I, in Escherichia coli. We then describe the preparation of SMA and the extraction and purification of rhodopsin molecules using SMA. For complete details on the use and execution of this protocol, please refer to Ueta et al. (2020).
AB - Microbial rhodopsins are photoreceptive membrane proteins showing various light-dependent biological activities. Styrene-maleic acid (SMA) copolymers spontaneously form nanoscale lipid particles containing membrane proteins and associated lipids without detergent, and can be used to characterize membrane molecules. Here, we provide a protocol to functionally express a thermally stable rhodopsin, Rubrobacter xylanophilus rhodopsin, and an unstable rhodopsin, Halobacterium salinarum sensory rhodopsin I, in Escherichia coli. We then describe the preparation of SMA and the extraction and purification of rhodopsin molecules using SMA. For complete details on the use and execution of this protocol, please refer to Ueta et al. (2020).
KW - Biotechnology and bioengineering
KW - Cell Membrane
KW - Protein Biochemistry
KW - Protein expression and purification
UR - http://www.scopus.com/inward/record.url?scp=85121266804&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85121266804&partnerID=8YFLogxK
U2 - 10.1016/j.xpro.2021.101046
DO - 10.1016/j.xpro.2021.101046
M3 - Article
C2 - 34984357
AN - SCOPUS:85121266804
SN - 2666-1667
VL - 3
JO - STAR Protocols
JF - STAR Protocols
IS - 1
M1 - 101046
ER -