Static and Dynamic Properties of Phospholipid Bilayer Nanodiscs

Nakano, Minoru; Fukuda, Masahiro; Kudo, Takayuki; Miyazaki, Masakazu; Wada, Yasushi; Matsuzaki, Naoya; Endo, Hitoshi; Handa, Tetsurou

doi:10.1021/ja9017013

Cited by 91 publications

(158 citation statements)

References 39 publications

(63 reference statements)

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“…This effect was previously reported by fluorescence spectroscopy on n‐AS (n‐(9‐anthroyloxy)stearic acid) inserted in large unilamellar vesicles (LUVs)35 and solid‐state 13 C NMR spectroscopy under magic angle spinning on nanodiscs 34. Whereas lipid signals are sharp for liposomes in the fluid phase, there is an increase of the C− 2 H bond line widths in nanodiscs at the same temperature.…”

Section: Resultssupporting

confidence: 62%

Lipid Internal Dynamics Probed in Nanodiscs

et al. 2017

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Nanodiscs offer a very promising tool to incorporate membrane proteins into native‐like lipid bilayers and an alternative to liposomes to maintain protein functions and protein–lipid interactions in a soluble nanoscale object. The activity of the incorporated membrane protein appears to be correlated to its dynamics in the lipid bilayer and by protein–lipid interactions. These two parameters depend on the lipid internal dynamics surrounded by the lipid‐encircling discoidal scaffold protein that might differ from more unrestricted lipid bilayers observed in vesicles or cellular extracts. A solid‐state NMR spectroscopy investigation of lipid internal dynamics and thermotropism in nanodiscs is reported. The gel‐to‐fluid phase transition is almost abolished for nanodiscs, which maintain lipid fluid properties for a large temperature range. The addition of cholesterol allows fine‐tuning of the internal bilayer dynamics by increasing chain ordering. Increased site‐specific order parameters along the acyl chain reflect a higher internal ordering in nanodiscs compared with liposomes at room temperature; this is induced by the scaffold protein, which restricts lipid diffusion in the nanodisc area.

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Section: Resultssupporting

confidence: 62%

Lipid Internal Dynamics Probed in Nanodiscs

et al. 2017

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“…Therefore spatial structures obtained in micelles are of the best quality but may be nonphysiological. Conversely, alternative membrane mimetics are also models of the lipid bilayer; lipid properties in nanodiscs and bicelles do not correspond to those in liposomes (51), and nanodiscs still cannot be used to solve the structures of transmembrane dimers due to their extremely large size. The bilayer thickness and state of the lipids are almost never known for a certain protein, so there always exists the possibility that incorrect choice of lipids in bicelles, nanodiscs, or liposomes can affect the state of the transmembrane protein, whereas micelles are much more flexible and can adopt their shape to the hydrophobic length of a transmembrane ␣-helix.…”

Section: Discussionmentioning

confidence: 99%

Structural Basis of p75 Transmembrane Domain Dimerization

Nadezhdin

Goncharuk

Mineev

et al. 2016

Journal of Biological Chemistry

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Dimerization of single span transmembrane receptors underlies their mechanism of activation. p75 neurotrophin receptor plays an important role in the nervous system, but the understanding of p75 activation mechanism is still incomplete. The transmembrane (TM) domain of p75 stabilizes the receptor dimers through a disulfide bond, essential for the NGF signaling. Here we solved by NMR the three-dimensional structure of the p75-TM-WT and the functionally inactive p75-TM-C257A dimers. Upon reconstitution in lipid micelles, p75-TM-WT forms the disulfide-linked dimers spontaneously. Under reducing conditions, p75-TM-WT is in a monomer-dimer equilibrium with the Cys 257 residue located on the dimer interface. In contrast, p75-TM-C257A forms dimers through the AXXXG motif on the opposite face of the ␣-helix. Biochemical and crosslinking experiments indicate that AXXXG motif is not on the dimer interface of p75-TM-WT, suggesting that the conformation of p75-TM-C257A may be not functionally relevant. However, rather than mediating p75 homodimerization, mutagenesis of the AXXXG motif reveals its functional role in the regulated intramembrane proteolysis of p75 catalyzed by the ␥-secretase complex. Our structural data provide an insight into the key role of the Cys 257 in stabilization of the weak transmembrane dimer in a conformation required for the NGF signaling.The neurotrophins (NTs) 6 are a family of neurotrophic factors that control multiple aspects of nervous system development and function. NTs interact with two distinct receptors, a cognate member of the Trk receptor tyrosine kinase family and the p75 neurotrophin receptor, a member of the tumor necrosis factor (TNF) receptor superfamily of death receptors (1). The most prominent biological function of p75 may be the induction of cell death, although it demonstrates several other activities, like survival, axonal growth, and cell migration (2-5).The precise mechanism as to how p75 transmits diverse signals in the normal or diseased nervous system remains elusive (3). Signal transduction by p75 is thought to proceed via the ligand-dependent recruitment and release of protein interactors to and from the receptor (6). Although different oligomer species have been recently described as the active receptor (7-10), several lines of evidence support the dimeric nature of p75 in neurotrophin signaling. NTs are homodimers in solution (11) that dimerize the extracellular domain of p75 upon binding as seen by in vivo cross-linking (12), by x-ray crystallography (13,14), and in solution (15). Engineered cysteine constructs activate p75 in the absence of NGF by inducing constitutive dimers (7), and the extracellular (16) and the intracellular domain (17, 18) form stable dimers in solution. In addition, the TM domain of p75 self-associates as measured by ToxCAT (8). All these data are supported by FRET (8,9) and by ␤-galactosidase protein-protein interaction assays (19), indicating that in the absence of NTs a significant fraction of p75 exists as preformed dimers on the plas...

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“…So far, various membrane proteins were reconstituted into nanodiscs (3)(4)(5). Due to their monodispersity, small angle x-ray (SAXS) and neutron scattering (SANS) could be used to confirm, not only the size and the disc-like bilayer structure of the nanodiscs (2,6) but also the physical properties of the bilayer patch (7,8). However, attempts to structurally characterize membrane proteins in nanodiscs are currently limited to SAXS studies of bacteriorhodopsin (9) and a human cytochrome P450 (CYP3A4) (10).…”

mentioning

confidence: 99%

Monitoring Shifts in the Conformation Equilibrium of the Membrane Protein Cytochrome P450 Reductase (POR) in Nanodiscs

Wadsäter

Laursen

Singha

et al. 2012

Journal of Biological Chemistry

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Background:Investigating the mechanism of NADPH-dependent conformational changes of POR in nanodiscs. Results:The conformational equilibrium of compact and extended POR, shifts toward the compact form (from 30 to 60%) upon reduction by NADPH. Conclusion:The NADPH-dependent conformational changes follow the "swinging model." Significance: This is the first time that the action of a membrane protein located in a lipid bilayer environment is probed by neutron reflectivity.

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Static and Dynamic Properties of Phospholipid Bilayer Nanodiscs

Cited by 91 publications

References 39 publications

Lipid Internal Dynamics Probed in Nanodiscs

Lipid Internal Dynamics Probed in Nanodiscs

Structural Basis of p75 Transmembrane Domain Dimerization

Monitoring Shifts in the Conformation Equilibrium of the Membrane Protein Cytochrome P450 Reductase (POR) in Nanodiscs

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