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 Aug 7)
PMID: 29937358
doi: 10.1016/j.str.2018.05.007 		Published in SASBDB:

SASDDB5 – 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) experimental SAS data
SASREF CV model
Sample: Lipid A export ATP-binding/permease protein MsbA dimer, 133 kDa Escherichia coli protein
Membrane scaffold protein 1D1 (deuterated, 75%) dimer, 49 kDa protein
1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl), 1 kDa Escherichia coli
Buffer: 30 mM Tris, 150 mM NaCl, pH: 7.5
Experiment: SANS data collected at D11, ILL on 2017 Mar 9
RgGuinier 4.0 nm
Dmax 13.0 nm
VolumePorod 189 nm3

SASDDC5 – MsbA in stealth nanodisc (SANS, 100% D2O) + 1 mM ADP

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) experimental SAS data
Lipid A export ATP-binding/permease protein MsbA Membrane scaffold protein 1D1 (deuterated, 75%) 1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl) Kratky plot
Sample: Lipid A export ATP-binding/permease protein MsbA dimer, 133 kDa Escherichia coli protein
Membrane scaffold protein 1D1 (deuterated, 75%) dimer, 49 kDa protein
1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl), 1 kDa Escherichia coli
Buffer: 30 mM Tris, 150 mM NaCl, 1 mM ADP, pH: 7.5
Experiment: SANS data collected at D11, ILL on 2017 Mar 9
RgGuinier 3.9 nm
Dmax 12.5 nm
VolumePorod 173 nm3

SASDDD5 – MsbA in stealth nanodisc (SAXS)

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) experimental SAS data
Lipid A export ATP-binding/permease protein MsbA Membrane scaffold protein 1D1 (deuterated, 75%) 1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl) Kratky plot
Sample: Lipid A export ATP-binding/permease protein MsbA dimer, 133 kDa Escherichia coli protein
Membrane scaffold protein 1D1 (deuterated, 75%) dimer, 49 kDa protein
1-palmitoyl-2-palmitoleoyl-sn-glycero-3-phosphocholine (deuteration: 78% head, 92% acyl), 1 kDa Escherichia coli
Buffer: 30 mM Tris, 150 mM NaCl, pH: 7.5
Experiment: SAXS data collected at EMBL P12, PETRA III on 2017 Sep 8
RgGuinier 4.8 nm
Dmax 16.0 nm
VolumePorod 607 nm3

SASDDE5 – NBD-MsbA (apo)

Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA experimental SAS data
Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA Kratky plot
Sample: Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA monomer, 27 kDa Escherichia coli protein
Buffer: 30 mM Tris, 150 mM NaCl, 0.5 mM TCEP, pH: 7.5
Experiment: SAXS data collected at EMBL P12, PETRA III on 2017 May 30
RgGuinier 2.2 nm
Dmax 7.3 nm
VolumePorod 47 nm3

SASDDF5 – NBD-MsbA (+1mM ADP)

Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA experimental SAS data
Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA Kratky plot
Sample: Nucleotide Binding Domain of Lipid A export ATP-binding/permease protein MsbA monomer, 27 kDa Escherichia coli protein
Buffer: 30 mM Tris, 150 mM NaCl, 0.5 mM TCEP, 1 mM ADP, pH: 7.5
Experiment: SAXS data collected at EMBL P12, PETRA III on 2017 May 30
RgGuinier 2.1 nm
Dmax 7.3 nm
VolumePorod 50 nm3