Activity and structure of EcoKMcrA.

Czapinska H, Kowalska M, Zagorskaite E, Manakova E, Slyvka A, Xu SY, Siksnys V, Sasnauskas G, Bochtler M, Nucleic Acids Res 46(18):9829-9841 (2018) Europe PMC

SASDJY2 – N-terminal domain of modification dependent EcoKMcrA restriction endonuclease

5-methylcytosine-specific restriction enzyme A (N-terminal domain)
MWI(0) 21 kDa
MWexpected 21 kDa
VPorod 29 nm3
log I(s) 1.24×103 1.24×102 1.24×101 1.24×100
5-methylcytosine-specific restriction enzyme A (N-terminal domain) small angle scattering data  s, nm-1
ln I(s)
5-methylcytosine-specific restriction enzyme A (N-terminal domain) Guinier plot ln 1.25×103 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
5-methylcytosine-specific restriction enzyme A (N-terminal domain) Kratky plot 1.104 0 3 sRg
p(r)
5-methylcytosine-specific restriction enzyme A (N-terminal domain) pair distance distribution function Rg: 2.2 nm 0 Dmax: 7.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
5-methylcytosine-specific restriction enzyme A (N-terminal domain) OTHER model

Synchrotron SAXS data from solutions of EcoKMcrA restriction endonuclease (N-terminal domain) in 20 mM Tris–HCl pH 7.5, 200 mM KCl, 0.1 mM EDTA, 0.01% (w/v) sodium azide, 1 mM DTT, were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1.3 and 3.5 mg/ml were measured at 20°C. 20 successive 0.050 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Prone to aggregation

5-methylcytosine-specific restriction enzyme A (N-terminal domain) (EcoKMcrA-N)
Mol. type   Protein
Organism   Escherichia coli K-12
Olig. state   Monomer
Mon. MW   20.6 kDa
 
UniProt   P24200 (1-175)
Sequence   FASTA