4XXJ

Crystal Structure of Escherichia coli-Expressed Halobacterium salinarum Bacteriorhodopsin in the Trimeric Form

Summary for 4XXJ

• Entry DOI: 10.2210/pdb4xxj/pdb
• Descriptor: Bacteriorhodopsin, EICOSANE, [(Z)-octadec-9-enyl] (2R)-2,3-bis(oxidanyl)propanoate, ... (4 entities in total)
• Functional Keywords: ion transport, proton pump
• Biological source: Halobacterium salinarum
• Cellular location: Cell membrane ; Multi-pass membrane protein : P02945
• Total number of polymer chains: 3
• Total formula weight: 98603.59

Authors

Bratanov, D.,Balandin, T.,Round, E.,Gushchin, I.,Gordeliy, V. (deposition date: 2015-01-30, release date: 2015-07-01, Last modification date: 2024-01-10)

Primary citation

Bratanov, D.,Balandin, T.,Round, E.,Shevchenko, V.,Gushchin, I.,Polovinkin, V.,Borshchevskiy, V.,Gordeliy, V.
An Approach to Heterologous Expression of Membrane Proteins. The Case of Bacteriorhodopsin.
Plos One, 10:e0128390-e0128390, 2015

PubMed 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.

PubMed: 26046789
DOI: 10.1371/journal.pone.0128390

Experimental method

X-RAY DIFFRACTION (1.9 Å)