Maltose-neopentyl glycol (MNG) amphiphiles for solubilization, stabilization and crystallization of membrane proteins

Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Nature Methods (Impact Factor: 32.07). 10/2010; 7(12):1003-8. DOI: 10.1038/nmeth.1526
Source: PubMed


The understanding of integral membrane protein (IMP) structure and function is hampered by the difficulty of handling these proteins. Aqueous solubilization, necessary for many types of biophysical analysis, generally requires a detergent to shield the large lipophilic surfaces of native IMPs. Many proteins remain difficult to study owing to a lack of suitable detergents. We introduce a class of amphiphiles, each built around a central quaternary carbon atom derived from neopentyl glycol, with hydrophilic groups derived from maltose. Representatives of this maltose-neopentyl glycol (MNG) amphiphile family show favorable behavior relative to conventional detergents, as manifested in multiple membrane protein systems, leading to enhanced structural stability and successful crystallization. MNG amphiphiles are promising tools for membrane protein science because of the ease with which they may be prepared and the facility with which their structures may be varied.

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Available from: Claus Juul Loland, Oct 07, 2015
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    • "A new promising class of detergents, the maltose-neopentyl glycols (MNGs), has been described recently. MNGs appear to be less destabilizing to MPs than DDM (Chae et al. 2010). One advantage of MNGs and, more generally, of detergents over APols and NDs is their ability to directly extract MPs from their native or host membranes. "
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    ABSTRACT: Solution-state nuclear magnetic resonance studies of membrane proteins are facilitated by the increased stability that trapping with amphipols confers to most of them as compared to detergent solutions. They have yielded information on the state of folding of the proteins, their areas of contact with the polymer, their dynamics, water accessibility, and the structure of protein-bound ligands. They benefit from the diversification of amphipol chemical structures and the availability of deuterated amphipols. The advantages and constraints of working with amphipols are discussed and compared to those associated with other non-conventional environments, such as bicelles and nanodiscs.
    Journal of Membrane Biology 03/2014; 247(9-10). DOI:10.1007/s00232-014-9654-z · 2.46 Impact Factor
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    • "The MNGs also confer greater stability on a range of different membrane protein molecules. Stability analysis revealed that the presence of the MNGs increased the T m of the β 2 AR + T4L compared to DDM [73]. Of all the detergent molecules tested MNG-3 (Fig. 5B) proved to be the best agent for both β 2 AR + T4L and a range of other integral membrane proteins. "
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    ABSTRACT: G protein-coupled receptors are integral membrane cell surface receptors with key roles in mediating the cellular responses to a wide range of biologically relevant molecules including hormones, neurotransmitters and importantly the majority of currently available drugs. The first high-resolution, X-ray crystallographic structure of a GPCR, that of rhodopsin, was obtained in 2000. It took a further seven years for the next structure, that of the β2 adrenergic receptor. Remarkably, at the time of writing, there have been an astonishing 18 further independent high-resolution GPCR structures published in the last five years (overall total of 68 structures in different conformations or bound to different ligands). Of particular note is the recent structure of the β2 adrenergic receptor in complex with its cognate heterotrimeric G-protein revealing for the first time molecular details of the interaction between a GPCR and the complete G-protein. Together these structures have provided unprecedented detail into the mechanism of action of these incredibly important proteins. This review describes several key methodological advances that have made such extraordinarily fast progress possible.
    Biochimica et Biophysica Acta 07/2013; 1828(11). DOI:10.1016/j.bbamem.2013.07.013 · 4.66 Impact Factor
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    • "One crucial step in establishing a purification protocol for a membrane protein is to select an appropriate detergent, which should not only be able to solubilize the membrane protein effectively but also to stabilize the protein and to preserve its biological activity. Recent work has shown that sevreal detergents, including FC-14, DDM, CHAPS, CHS, OG, Brij-35 and some detergent mixtures, may be good for maintaining the structure and function of integral membrane proteins [22], [24], [25], [37], [38], [39]. In this work, several detergents were therefore used to screen for better solubilization of hCCR3. "
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    ABSTRACT: Human chemokine receptor CCR3 (hCCR3) belongs to the G protein-coupled receptors (GPCRs) superfamily of membrane proteins and plays major roles in allergic diseases and angiogenesis. In order to study the structural and functional mechanism of hCCR3, it is essential to produce pure protein with biological functions on a milligram scale. Here we report the expression of hCCR3 gene in a tetracycline-inducible stable mammalian cell line. A cell clone with high hCCR3 expression was selected from 46 stably transfected cell clones and from this cell line pure hCCR3 on a milligram scale was obtained after two-step purification. Circular dichroism spectrum with a characteristic shape and magnitude for α-helix indicated proper folding of hCCR3 after purification. The biological activity of purified hCCR3 was verified by its high binding affinity with its endogenous ligands CCL11 and CCL24, with K D in the range of 10(-8) M to 10(-6) M.
    PLoS ONE 06/2013; 8(6):e65500. DOI:10.1371/journal.pone.0065500 · 3.23 Impact Factor
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