| MWI(0) | 145 | kDa |
| MWexpected | 183 | kDa |
| VPorod | 189 | nm3 |
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log I(s)
8.90×10-1
8.90×10-2
8.90×10-3
8.90×10-4
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s, nm-1
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Conformational States of ABC Transporter MsbA in a Lipid Environment Investigated by Small-Angle Scattering Using Stealth Carrier Nanodiscs.
Josts I, Nitsche J, Maric S, Mertens HD, Moulin M, Haertlein M, Prevost S, Svergun DI, Busch S, Forsyth VT, Tidow H, Structure 26(8):1072-1079.e4 (2018) Europe PMCSASDDB5 – MsbA in stealth nanodisc (SANS, 100% D2O)
Lipid A export ATP-binding/permease protein MsbAMembrane scaffold protein 1D1 (deuterated, 75%)
1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl)
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Fits and models
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SANS
data from solutions of
MsbA in stealth nanodisc (SANS, 100% D2O)
in
30 mM Tris, 150 mM NaCl, pH 7.5
were collected
using a CERCA detector
at a wavelength of λ = 0.46 nm
(I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).
Solute concentrations ranging between 1 and 6 mg/ml were measured
at 10°C.
Two successive
4200 second frames were collected.
The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
MsbA ABC transporter protein encapsulated in a partially deuterated "stealth-nanodisc": (sND). Disc components (MSP1D1 scaffold protein - 75% deuterated, and POPC lipid - heads/tails 78% and 92% deuterated) are effectively matched out at 100% D2O. Thus the expected molecular weight should correspond to that of the MsbA protein (~132 kDa). Data measured in two configurations: 1.5 m / 1.5 m and 8 m / 8 m (collimation length / sample-detector distances) with a fixed neutron wavelength (λ) of 5 Å. Data is merged from these two configurations. |
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