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

SASDRX3 – Trypsin + Aprotinin: Complex equilibrium

Serine protease 1
Pancreatic trypsin inhibitor
MWexperimental 19 kDa
MWexpected 37 kDa
VPorod 31 nm3
log I(s) 2.84×101 2.84×100 2.84×10-1 2.84×10-2
Serine protease 1 Pancreatic trypsin inhibitor small angle scattering data  s, nm-1
ln I(s)
Serine protease 1 Pancreatic trypsin inhibitor Guinier plot ln 2.84×101 Rg: 1.9 nm 0 (1.9 nm)-2 s2
(sRg)2I(s)/I(0)
Serine protease 1 Pancreatic trypsin inhibitor Kratky plot 1.104 0 3 sRg
p(r)
Serine protease 1 Pancreatic trypsin inhibitor pair distance distribution function Rg: 1.9 nm 0 Dmax: 5.3 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Trypsin + Aprotinin: Complex equilibrium in 20 mM TRIS, 40 mM KCl, 20 mM CaCl2, pH 7 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). One solute concentration of 5.33 mg/ml was 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.

Aprotinin (1.73 mg/mL) and trypsin (4 mg/mL) were combined and static data point was taken minutes later. Molecular weight determined with Vc

Serine protease 1 (Trypsin)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   25.8 kDa
 
UniProt   P00760 (1-246)
Sequence   FASTA
 
Pancreatic trypsin inhibitor (Aprotinin)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   10.9 kDa
 
UniProt   P00974 (1-100)
Sequence   FASTA