Structural Insights on Dynamic behavior of PDI in solution environment through SAXS technique

Chandra Sekar.

SASDH52 – Ribonuclease A (Type III-A) in a solution environment at pH 7.5

Ribonuclease pancreatic
MWI(0) 17 kDa
MWexpected 14 kDa
VPorod 20 nm3
log I(s) 7.61×100 7.61×10-1 7.61×10-2 7.61×10-3
Ribonuclease pancreatic small angle scattering data  s, nm-1
ln I(s)
Ribonuclease pancreatic Guinier plot ln 7.61×100 Rg: 1.9 nm 0 (1.9 nm)-2 s2
(sRg)2I(s)/I(0)
Ribonuclease pancreatic Kratky plot 1.104 0 3 sRg
p(r)
Ribonuclease pancreatic pair distance distribution function Rg: 1.9 nm 0 Dmax: 7.2 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Ribonuclease pancreatic DAMMIF model
Ribonuclease pancreatic DAMMIF model

log I(s)
 s, nm-1
Ribonuclease pancreatic DAMMIF model
Ribonuclease pancreatic DAMMIF model

log I(s)
 s, nm-1
Ribonuclease pancreatic DAMFILT model

Synchrotron SAXS data from solutions of Ribonuclease A (Type III-A) in a solution environment at pH 7.5 in 50 mM Tris-HCL, 150 mM NaCl, pH 7.5 were collected on the BioCAT 18ID beam line at the Advanced Photon Source (APS), Argonne National Laboratory (Lemont, IL, USA) using a Pilatus3 X 1M detector at a sample-detector distance of 3.5 m and at a wavelength of λ = 0.1033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2.5 and 10 mg/ml were measured at 10°C. Eight successive 1 second frames were collected. Using the automated data processing pipeline at the beam line, the data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve. Subsequent data processing and three-dimensional modeling were performed using the ATSAS [2.8.3] package.

Ribonuclease A (Type III-A) was purchased from SIGMA-ALDRICH in the form of lyophilized powder, dissolved with suspension medium and used directly without any further purification.

Ribonuclease pancreatic (RNase-A)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   13.7 kDa
 
UniProt   P61823 (27-150)
Sequence   FASTA