A Tail-Based Mechanism Drives Nucleosome Demethylation by the LSD2/NPAC Multimeric Complex.

Marabelli C, Marrocco B, Pilotto S, Chittori S, Picaud S, Marchese S, Ciossani G, Forneris F, Filippakopoulos P, Schoehn G, Rhodes D, Subramaniam S, Mattevi A, Cell Rep 27(2):387-399.e7 (2019) Europe PMC

SASDFX3 – Semi-synthetic nucleosome core particle (NCP)

Histone H3
Histone H4
Histone H2a
Histone H2b
147bp 601 Widom sequence
MWexperimental 200 kDa
MWexpected 153 kDa
VPorod 353 nm3
log I(s) 9.12×101 9.12×100 9.12×10-1 9.12×10-2
Histone H3 Histone H4 Histone H2a Histone H2b 147bp 601 Widom sequence small angle scattering data  s, nm-1
ln I(s)
Histone H3 Histone H4 Histone H2a Histone H2b 147bp 601 Widom sequence Guinier plot ln 9.13×101 Rg: 4.4 nm 0 (4.4 nm)-2 s2
(sRg)2I(s)/I(0)
Histone H3 Histone H4 Histone H2a Histone H2b 147bp 601 Widom sequence Kratky plot 1.104 0 3 sRg
p(r)
Histone H3 Histone H4 Histone H2a Histone H2b 147bp 601 Widom sequence pair distance distribution function Rg: 4.3 nm 0 Dmax: 12.8 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of a semi-synthetic nucleosome core particle (NCP) in 15 mM HEPES, 200 mM NaCl, pH 7.3 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 20 °C using the following parameters: Column: Wyatt WTC-030N5; Flow rate: 0.25 mL/min; Sample injection concentration: 2.5 mg/mL; Injection volume: 50 μL. The data were collected through the SEC peak of the sample as a series of 243 x 1 second exposures. Each unsubtracted data frame was normalised to the intensity of the transmitted beam and radially averaged and the scattering of an appropriate solvent-blank was subtracted. The resulting subtracted frames were scaled and averaged to generate the final SAXS profile displayed in this entry.

Histone H3 (H3)
Mol. type   Protein
Organism   Xenopus laevis
Olig. state   Dimer
Mon. MW   15.3 kDa
Sequence   FASTA
 
Histone H4 (H4)
Mol. type   Protein
Organism   Xenopus laevis
Olig. state   Dimer
Mon. MW   11.2 kDa
 
UniProt   P62799 (2-103)
Sequence   FASTA
 
Histone H2a (H2a)
Mol. type   Protein
Organism   Xenopus laevis
Olig. state   Dimer
Mon. MW   14.0 kDa
 
UniProt   Q6AZJ8 (2-130)
Sequence   FASTA
 
Histone H2b (H2b)
Mol. type   Protein
Organism   Xenopus laevis
Olig. state   Dimer
Mon. MW   13.5 kDa
 
UniProt   Q92130 (5-126)
Sequence   FASTA
 
147bp 601 Widom sequence (601-147bp)
Mol. type   DNA
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   45.2 kDa
Sequence   FASTA