Rational thermostabilisation of four-helix bundle dimeric de novo proteins.

Irumagawa S, Kobayashi K, Saito Y, Miyata T, Umetsu M, Kameda T, Arai R, Sci Rep 11(1):7526 (2021) Europe PMC

SASDKQ8 – Rationally optimised SUWA (ROSA) dimer

Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer
MWI(0) 25 kDa
MWexpected 25 kDa
VPorod 30 nm3
log I(s) 4.82×10-2 4.82×10-3 4.82×10-4 4.82×10-5
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer small angle scattering data  s, nm-1
ln I(s)
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer Guinier plot ln 4.82×10-2 Rg: 2.5 nm 0 (2.5 nm)-2 s2
(sRg)2I(s)/I(0)
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer Kratky plot 1.104 0 3 sRg
p(r)
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer pair distance distribution function Rg: 2.6 nm 0 Dmax: 10.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer DAMMIN model

log I(s)
 s, nm-1
Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer GROMACS model

Synchrotron SAXS data from solutions of Rationally optimised SUWA (ROSA) dimer in 20 mM HEPES, 150 mM NaCl, 5% glycerol,, pH 7.5 were collected on the BL-10C beam line at the Photon Factory (PF), High Energy Accelerator Research Organization (KEK) storage ring (Tsukuba, Japan) using a Pilatus3 2M detector at a sample-detector distance of 1.1 m and at a wavelength of λ = 0.13 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 3.22 mg/ml was measured at 20°C. 30 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.

Rationally optimised SUWA mutant N22E/H86K (ROSA) dimer (ROSA dimer)
Mol. type   Protein
Organism   Artificial protein (de novo protein)
Olig. state   Dimer
Mon. MW   12.6 kDa
Sequence   FASTA
 
PDB code   6KOS