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Synchrotron SAXS data from solutions of sweet potato beta-amylase 5 in 20 mM HEPES, 150 mM NaCl, and 0.2 mM TCEP, pH 7.3 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS) storage ring (Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).
IbBAM5 from Sigma-Aldrich was resuspended to 7 mg/mL in 20 mM HEPES (pH 7.3), 150 mM NaCl, and 0.2 mM TCEP and separated by HiLoad 16/60 Superdex 200 column equilibrated with SEC buffer (20 mM HEPES, pH 7.3, 150 mM NaCl, and 0.2 mM TCEP.). Purified IbBAM5 was diluted to concentrations between 1 and 10 mg mL-1 in 20 mM HEPES, pH 7.3, 150 mM NaCl, and 0.2 mM TCEP and flash frozen in the plate using liquid nitrogen. The sample plate was shipped overnight on dry ice to the Advanced Light Source at Lawrence Berkeley National Laboratory. Prior to data collection (date of collection: 12/07/2020), the plate was spun at 3700 rev min-1 for 10 minutes by beam line staff. Scattering data on the samples and controls were collected every 0.3 seconds for a total of 10 seconds resulting in 33 frames of data per sample. The beam energy was 11 keV, and the detector was 2 meters from the sample holder. Samples were kept at 10 °C during collection. Buffer scattering was subtracted from sample scattering in RAW (version 2.0.3) using SAXS FrameSlice (version 1.4.13) as a guide to determine which frames were used during buffer subtraction (Hopkins et al., 2017).
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s, nm-1
