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

SASDRJ4 – GAC rRNA + L11 Protein: SEC-SAXS final

58 nucleotide RNA L11-binding domain from E. coli 23S rRNA
50S ribosomal protein L11
MWexperimental 31 kDa
MWexpected 34 kDa
VPorod 48 nm3
log I(s) 4.82×10-2 4.82×10-3 4.82×10-4 4.82×10-5
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA 50S ribosomal protein L11 small angle scattering data  s, nm-1
ln I(s)
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA 50S ribosomal protein L11 Guinier plot ln 4.82×10-2 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
58 nucleotide RNA L11-binding domain from E. coli 23S rRNA 50S ribosomal protein L11 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 GAC rRNA + L11 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). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A sample at 3.2 mg/ml was injected onto a column 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.

SEC-SAXS profile of the final GAC rRNA + L11 Protein complex. Under standard equilibrium conditions, the combination of GAC rRNA + L11 protein forms a mixed state. Molecular Weight determined by Vc. SEC column = UNKNOWN. SEC sample injection volume = UNKNOWN. SEC flow rate = UNKNOWN.

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
 
50S ribosomal protein L11 (L11 protein)
Mol. type   Protein
Organism   Thermus thermophilus
Olig. state   Monomer
Mon. MW   15.5 kDa
 
UniProt   P36238 (1-147)
Sequence   FASTA