|
Synchrotron SAXS
data from solutions of
Tumbleweed protein composed of synthetic constructs TW1 and TW2 linked covalently via SpyTag/SpyCatcher
in
10 mM HEPES, 150 mM NaCl, 5mM MgCl2, pH 7.4
were collected
on the
SAXS/WAXS beam line
at the Australian Synchrotron storage ring
(Melbourne, Australia)
using a Pilatus3 S 2M detector
at a sample-detector distance of 2.7 m and
at a wavelength of λ = 0.10332 nm
(I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).
One solute concentration of 10.00 mg/ml was measured
at 20°C.
10 successive
1 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.
TW is a protein formed by covalent linkage of the the proteins TW1 and TW2 via a SpyTag/SpyCatcher. SAXS data was collected at the SAXS/WAXS beamline at the Australian Synchrotron, with Pilatus 2M detector at a SDD of 2680mm, with an incident intensity of ~8 x 10^12 ph/s. A SEC-SAXS setup with sheath flow was used. The running buffer was 10 mM HEPES pH 7.4, 150 mM NaCl, 5mM MgCl2, and 50uL of sample at ~10mg/mL was injected onto a Superdex 200 Increase 5/150 GL running at 0.2mL/min. Buffer scattering frames taken immediately before the void fraction. Sample scattering frames taken at the elution peak over a region of stable Rg using CHROMIX. Data were fit by a 2-state model using MultiFoXS.
|
|
Tumbleweed
(TW)
|
| Mol. type |
|
Protein |
| Organism |
|
synthetic construct |
| Olig. state |
|
Monomer |
| Mon. MW |
|
132.8 kDa |
| Sequence |
|
FASTA |
| |
|
s, nm-1
