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

SASDR54 – Kenics GAC rRNA + Mg: Time-Resolved 631 ms

58 nucleotide RNA L11-binding domain from E. coli 23S rRNA
MWexperimental 18 kDa
MWexpected 19 kDa
VPorod 26 nm3
log I(s) 1.25×101 1.25×100 1.25×10-1 1.25×10-2
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 1.25×101 Rg: 2.3 nm 0 (2.3 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: 9.8 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Kenics GAC rRNA + Mg: Time-Resolved 631 ms in 10 mM Na-MOPSO, 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 1.1 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 (60-80 micromolar) and MgCl2 (10 mM) were mixed together with a Kenics-style, chaotic advection mixer. Molecular weight determined by Vc.

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