Using stable MutS dimers and tetramers to quantitatively analyze DNA mismatch recognition and sliding clamp formation.

Groothuizen FS, Fish A, Petoukhov MV, Reumer A, Manelyte L, Winterwerp HH, Marinus MG, Lebbink JH, Svergun DI, Friedhoff P, Sixma TK, Nucleic Acids Res 41(17):8166-81 (2013) Europe PMC

SASDAN3 – MutS dimer

DNA mismatch repair protein MutS
MWI(0) 160 kDa
MWexpected 191 kDa
VPorod 307 nm3
log I(s) 1.05×104 1.05×103 1.05×102 1.05×101
DNA mismatch repair protein MutS small angle scattering data  s, nm-1
ln I(s)
DNA mismatch repair protein MutS Guinier plot ln 1.05×104 Rg: 4.7 nm 0 (4.7 nm)-2 s2
DNA mismatch repair protein MutS Kratky plot 1.104 0 3 sRg
DNA mismatch repair protein MutS pair distance distribution function Rg: 4.6 nm 0 Dmax: 15.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
DNA mismatch repair protein MutS DAMMIF model

Synchrotron SAXS data from solutions of MutS dimer in 50 mM HEPES 50 mM KCl, pH 7.5 were collected on the EMBL P12 camera at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.12 nm , the range of momentum transfer 0.076 < s < 4.423 nm-1 was covered (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.37 mg/ml was measured at 10°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

DNA mismatch repair protein MutS (MutS)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Dimer
Mon. MW   95.3 kDa
UniProt   P23909
Sequence   FASTA