Insights on the dynamic behavior of protein disulfide isomerase in the solution environment through the SAXS technique

Sanyasi C, Balakrishnan S, Chinnasamy T, Venugopalan N, Kandavelu P, Batra-Safferling R, Muthuvel S, In Silico Pharmacology 12(1) (2024) DOI

SASDH42 – Bovine Pancreatic Trypsin Inhibitor (BPTI) in solution environment at pH 7.5

Pancreatic trypsin inhibitor
MWI(0) 22 kDa
MWexpected 13 kDa
VPorod 6 nm3
log I(s) 8.55×100 8.55×10-1 8.55×10-2 8.55×10-3
Pancreatic trypsin inhibitor small angle scattering data  s, nm-1
ln I(s)
Pancreatic trypsin inhibitor Guinier plot ln 8.55×100 Rg: 1.4 nm 0 (1.4 nm)-2 s2
(sRg)2I(s)/I(0)
Pancreatic trypsin inhibitor Kratky plot 1.104 0 3 sRg
p(r)
Pancreatic trypsin inhibitor pair distance distribution function Rg: 1.4 nm 0 Dmax: 4.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Pancreatic trypsin inhibitor DAMMIF model
Pancreatic trypsin inhibitor DAMMIF model

log I(s)
 s, nm-1
Pancreatic trypsin inhibitor DAMMIF model
Pancreatic trypsin inhibitor DAMMIF model

log I(s)
 s, nm-1
Pancreatic trypsin inhibitor DAMFILT model

Synchrotron SAXS data from solutions of Bovine Pancreatic Trypsin Inhibitor (BPTI) in 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.

Bovine Pancreatic Trypsin Inhibitor (BPTI) was purchased from SIGMA-ALDRICH in the form of lyophilized powder, dissolved with suspension medium and used directly without any further purification. CAUTION! Sample likely aggregated.

Pancreatic trypsin inhibitor (BPTI)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Dimer
Mon. MW   6.5 kDa
 
UniProt   P00974 (36-93)
Sequence   FASTA