Chaotic advection mixer for capturing transient states of diverse biological macromolecular systems with time-resolved small-angle X-ray scattering

Zielinski K, Katz A, Calvey G, Pabit S, Milano S, Aplin C, San Emeterio J, Cerione R, Pollack L, IUCrJ 10(3):363-375 (2023) DOI

SASDRP4 – Diffusive GAC rRNA + Mg: Time-Resolved 1000 ms

58 nucleotide RNA L11-binding domain from E. coli 23S rRNA
MWexperimental 20 kDa
MWexpected 19 kDa
VPorod 30 nm3
log I(s) 8.72×10-6 8.72×10-7 8.72×10-8 8.72×10-9
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA small angle scattering data  s, nm-1
ln I(s)
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA Guinier plot ln 8.72×10-6 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA Kratky plot 1.104 0 √3 sRg
Dmax: 13 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Diffusive GAC rRNA + Mg: Time-Resolved 1000 ms in 10 mM sodium cacodylate, 100 mM KCl, pH 6.5 were collected on the G1 beam line at the Cornell High Energy Synchrotron Source (CHESS) storage ring (Ithaca, NY, USA) using a Pilatus 100K detector at a wavelength of λ = 0.10972 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1.2 and 1.5 mg/ml were measured at 20°C. 20 successive 5 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.

GAC rRNA (75-100 micromolar) and MgCl2 (20 mM) were rapidly mixed together with a diffusive mixer. Molecular weight determined by Vc. Data published in Welty, R., Pabit, S. A., Katz, A. M., Calvey, G. D., Pollack, L. & Hall, K. B. (2018). RNA 24, 1828–1838. https://doi.org/10.1261/rna.068361.118

58 nucleotide RNA L11-binding domain from E. coli 23S rRNA (GAC rRNA)
Mol. type   RNA
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   18.8 kDa
Sequence   FASTA