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Review
. 2012 Nov 14;112(11):6054-74.
doi: 10.1021/cr300061w. Epub 2012 Aug 24.

The magic of bicelles lights up membrane protein structure

Ulrich H N Dürr  1 , Melissa Gildenberg, Ayyalusamy Ramamoorthy
Affiliations
Free PMC article
Review

The magic of bicelles lights up membrane protein structure

Ulrich H N Dürr et al. Chem Rev. .
Free PMC article
No abstract available

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Figures

Figure 1
Figure 1
A schematic overview of the use of lipid bicelles in the study of structure and dynamics of membrane proteins using NMR spectroscopy.
Figure 2
Figure 2
Schematic models for the morphology of bicellar phases with increasing detergent content: multilamellar vesicles (A), with toroidal pores lined up by detergents (B), extended lamellae showing magnetic-alignment (C), chiral nematic “worm-like” ribbons, also magnetically alignable (D), flat disk-like aggregates tumbling isotropically (E), and detergent micelles (F).
Figure 3
Figure 3
Proteins can be studied in numerous ways employing bicelles. Membrane proteins can be macroscopically oriented using magnetically aligned bicelles (A). Magnetically aligned bicelles can also be used to give a residual preferential orientation to globular proteins (B). Membrane proteins can be studied in isotropically tumbling bicelles (C). The combination of isotropic bicelles and globular proteins can be used to study membrane binding (D).
Figure 4
Figure 4
Preparation protocols for a bicelle containing membrane proteins can be chosen from a variety of possible pathways. Red, dark green, and light green colors denote protein, lipid bilayer, and detergent, respectively.
Figure 5
Figure 5
Schematic of the topology of full-length cytochrome b5 (cyt b5, yellow). The protein consists of an α-helical transmembrane domain, a highly flexible linker region, and a globular domain that carries a prosthetic heme molecule. Also shown is the bicellar environment used to macroscopically align cyt b5 with respect to the external magnetic field, B0.
Figure 6
Figure 6
Proton-decoupled 31P NMR chemical shift spectra of different bicelle preparations used in the study of cytochrome b5. Phosphorus-31 NMR spectra report directly on the quality of bicelle alignment. (A) Pure q = 3.5 DMPC/DHPC bicelles. Bicelles in the presence of one cytochrome b5 molecule per 86 (B), 170 (C), and 212 (D) molecules of DMPC. (E) Bicelles in the presence of both cytochrome b5 and cytochrome P450. The resonance observed at approximately 0 ppm originates from phosphate buffer.
Figure 7
Figure 7
Structural observations on full-length cytochrome b5 using solid-state 15N NMR spectroscopy. (Top) Molecular model of cytochrome b5 in a lipid bilayer. (Middle) One-dimensional 15N NMR spectra and (bottom) two-dimensional HIMSELF spectrum of uniformly 15N-labeled cytochrome b5 embedded in DMPC/DHPC q = 3.5 bicelles.
Figure 8
Figure 8
15N Chemical shift spectra of oriented bicelles containing cytochrome b5 in RampCP and RINEPT (A) and DREPT (B) experiments. Refocusing delay times vary as indicated in (B). Short refocusing periods are sufficient for the production of peaks from cytochrome b5’s rigid domain while longer delay times are necessary for the detection of soluble domain resonances. (C, D) 2D spectra obtained using the RINEPT sequence with refocusing delays of 1 ms and 80 μs, respectively. Reprinted with permission from ref (286). Copyright 2010 American Chemical Society.
Figure 9
Figure 9
Spectra showing the 15N isotropic chemical shift of 15N-labeled rabbit cyt b5 in q = 3.5 DMPC/DHPC bicelles (A). Comparison of RampCP (a) and RINEPT (b) polarization transfer schemes, where RampCP shows arginine and lysine side chain resonances. 2D 13C chemical shift correlation spectra of 13C,15N-labeled rabbit cyt b5 in bicelles (B). CTUC COSY (a) and DARR (b) spectra are pictured, with amino acid cross peaks labeled in (a). Collage of parts of two figures reprinted with permission from ref (283). Copyright 2008 John Wiley & Sons, Inc.
Figure 10
Figure 10
Order parameter profiles of pure DMPC bilayers (open symbols) and DMPC bilayers in the presence of 10 wt % myelin basic protein (filled), lines are meant to guide the eye. On top, a DMPC molecular scheme gives the employed site naming scheme.
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References

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