Engineered variants provide new insight into the structural properties important for activity of the highly dynamic, trimeric protein disulfide isomerase ScsC from Proteus mirabilis.

Furlong EJ, Kurth F, Premkumar L, Whitten AE, Martin JL, Acta Crystallogr D Struct Biol 75(Pt 3):296-307 (2019) Europe PMC

SASDEQ4 – Deletion mutant of the suppressor of copper sensitivity C protein, PmScsC, with concentration series data

Deletion mutant of PmScsC
MWexperimental 44 kDa
MWexpected 23 kDa
VPorod 54 nm3
log I(s) 2.42×10-1 2.42×10-2 2.42×10-3 2.42×10-4
Deletion mutant of PmScsC small angle scattering data  s, nm-1
ln I(s)
Deletion mutant of PmScsC Guinier plot ln 2.42×10-1 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Deletion mutant of PmScsC Kratky plot 1.104 0 √3 sRg
p(r)
Deletion mutant of PmScsC pair distance distribution function Rg: 2.6 nm 0 Dmax: 9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Deletion mutant of PmScsC PDB (PROTEIN DATA BANK) model
X-ray synchrotron radiation scattering data from solutions of a deletion mutant of Suppressor of Copper Sensitivity C protein (ScsC) protein from Proteus mirabilis in 10 mM HEPES 150mM NaCl, pH 7.4 were collected on the SAXS/WAXS beam line of the Australian Synchrotron (Melbourne, Australia) using a 2D Photon counting Pilatus 1M-W pixel detector (s = 4π sin θ/λ, where 2θ is the scattering angle). Thirty eight successive 1 second frames were collected across solute concentrations of 0.1-10.0 mg/ml at a sample temperature of 10°C. The data were normalized to the intensity of the transmitted beam and radially averaged and the scattering of the solvent-blank was subtracted. The SAXS data displayed this entry was derived from a 10.0 mg/ml sample and show that a mixture of monomer, dimer and trimer is present in solution. A linear combination of monomer, dimer and trimer are fit to the scattering curve, and the final concentration of each species in solution was determined to be: 122.70 µM (monomer); 73.15 µM (dimer); 12.29 µM (trimer). Together, with data collected at other concentration (available to download as a zip archive), it was estimated that K1 (monomer <-> dimer) was 205.81 µM, and K2 (monomer + dimer <-> trimer) 729.98 µM.

Deletion mutant of PmScsC (PmScsC&Delta;L)
Mol. type   Protein
Organism   Proteus mirabilis
Olig. state   Unknown
Mon. MW   23.4 kDa
 
UniProt   A0A1Z1SYD5 (22-243)
Sequence   FASTA
 
PDB ID   4XVW
 
PDB ID   6MHH