| MWexperimental | 136 | kDa |
| MWexpected | 168 | kDa |
| VPorod | 217 | nm3 |
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log I(s)
9.20×10-2
9.20×10-3
9.20×10-4
9.20×10-5
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s, nm-1
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Structural basis for activation of plasma-membrane Ca2+-ATPase by calmodulin.
Nitsche J, Josts I, Heidemann J, Mertens HD, Maric S, Moulin M, Haertlein M, Busch S, Forsyth VT, Svergun DI, Uetrecht C, Tidow H, Commun Biol 1:206 (2018) Europe PMCSASDET4 – ACA8 complex with Calmodulin (75% deuterated) in stealth nanodisc (SANS, 100% D2O)
Membrane scaffold protein 1D1 (deuterated, 75%)1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl)
Calcium-transporting ATPase 8, plasma membrane-type
Calmodulin-7 (deuterated 75%)
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Fits and models
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Rg, nm
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SANS
data from solutions of
ACA8 complex with Calmodulin (75% deuterated) in stealth nanodisc (SANS, 100% D2O)
in
30 mM Tris, 150 mM NaCl, 1mM MgCl2, 1 mM CaCl2, pH 7.5
were collected
using a MLZ detector
at a sample-detector distance of 5.5 m and
at a wavelength of λ = 5 nm
(I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).
Solute concentrations ranging between 3.2 and 3.2 mg/ml were measured
at 10°C.
One
7200 second frame was collected.
The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
ACA8 protein complex with Calmodulin (75% deuterated) encapsulated in a partially deuterated "stealth-nanodisc": (sND). The Calmodulin and 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 ACA8 protein alone (~118 kDa). Data measured using a single instrument configuration: 5.5 m (sample-detector distances) with a fixed neutron wavelength (λ) of 5 Å. |
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