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

SASDRG3 – Tissue Transglutaminase + Ca: Time-resolved 63 ms

Protein-glutamine gamma-glutamyltransferase 2
MWexperimental 117 kDa
MWexpected 77 kDa
VPorod 155 nm3
log I(s) 9.69×10-6 9.69×10-7 9.69×10-8 9.69×10-9
Protein-glutamine gamma-glutamyltransferase 2 small angle scattering data  s, nm-1
ln I(s)
Protein-glutamine gamma-glutamyltransferase 2 Guinier plot ln 9.69×10-6 Rg: 4.1 nm 0 (4.1 nm)-2 s2
(sRg)2I(s)/I(0)
Protein-glutamine gamma-glutamyltransferase 2 Kratky plot 1.104 0 3 sRg
Dmax: 18 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Tissue Transglutaminase + Ca: Time-resolved 63 ms in 20 mM HEPES, 100 mM NaCl, 10% glycerol, 1 mM DTT, pH 7.5 were collected on the ID7A1 BioSAXS / HP-Bio Beamline beam line at the Cornell High Energy Synchrotron Source (CHESS) storage ring (Ithaca, NY, USA) using a Eiger 4M detector at a wavelength of λ = 0.10972 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 1.00 mg/ml was measured at 20°C. 15 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.

2 mg/mL ttG + 4 mM CaCl2 were rapidly mixed together with a Kenics style chaotic advection mixer. Final concentration during observation is 1 mg/mL ttG + 2 mM CaCl2. Molecular weight from Vc.

Protein-glutamine gamma-glutamyltransferase 2 (TG2)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Unknown
Mon. MW   77.3 kDa
 
UniProt   P21980 (1-687)
Sequence   FASTA