Abstract
The Saccharomyces (S.) cerevisiae mitochondrial respiratory supercomplex factor 2 (Rcf2) plays a role in assembly of supercomplexes composed of cytochrome bc1 (complex III) and cytochrome c oxidase (complex IV). The Rcf2 protein was here expressed in E. coli, refolded and reconstituted into dodecylphosphocholine (DPC) micelles. The structural properties of Rcf2 were studied by solution NMR and near complete backbone assignment of Rcf2 was achieved. The secondary structure of Rcf2 contains seven helices, of which five are putative transmembrane (TM) helices, including, unexpectedly, a region formed by a charged 20-residue helix at the C-terminus. Further studies demonstrated that Rcf2 forms a dimer and the charged TM helix is involved in this dimer formation. Our results provide a basis for understanding the role of this assembly/regulatory factor in supercomplex formation and function.
PMID: 29240987 [PubMed - as supplied by publisher]
Dodecyl-?-melibioside Detergent Micelles asa Medium for Membrane Proteins
Dodecyl-?-melibioside Detergent Micelles asa Medium for Membrane Proteins
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.7b00810/20171006/images/medium/bi-2017-00810n_0005.gif
Biochemistry
DOI: 10.1021/acs.biochem.7b00810
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[NMR paper] Expression, purification and reconstitution of the C-terminal transmembrane domain of scavenger receptor BI into detergent micelles for NMR analysis.
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[NMR paper] Structural Basis for TatA Oligomerization: An NMR Study of Escherichia coli TatA Dimeric Structure.
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[NMR paper] Solution-NMR Characterization of Outer-Membrane Protein A from E. coli in Lipid Bilayer Nanodiscs and Detergent Micelles.
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[NMR paper] Solution NMR spectroscopic characterization of human VDAC-2 in detergent micelles and lipid bilayer nanodiscs.
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Micro-coil NMR to monitor optimization of the reconstitution conditions for the integral membrane protein OmpW in detergent micelles
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One of the major obstacles to membrane protein (MP) structural studies is the destabilizing effect of detergents. Amphipols (APols) are short amphipathic polymers that can substitute for detergents to keep MPs water-soluble under mild conditions. In the present work, we have explored the...
NMR study of Rcf2 reveals an unusual dimeric topology in detergent micelles.
Linear Mode
