How the central domain of dystrophin acts to bridge F-actin to sarcolemmal lipids.

Mias-Lucquin D, Dos Santos Morais R, Chéron A, Lagarrigue M, Winder SJ, Chenuel T, Pérez J, Appavou MS, Martel A, Alviset G, Le Rumeur E, Combet S, Hubert JF, Delalande O, J Struct Biol :107411 (2019) Europe PMC

SASDFW4 – Conformation of R8-15 human dystrophin fragment

Human dystrophin central domain R8-15 fragment
MWexperimental 88 kDa
MWexpected 100 kDa
log I(s) 3.27×10-2 3.27×10-3 3.27×10-4 3.27×10-5
Human dystrophin central domain R8-15 fragment small angle scattering data  s, nm-1
ln I(s)
Human dystrophin central domain R8-15 fragment Guinier plot ln 3.27×10-2 Rg: 10.1 nm 0 (10.1 nm)-2 s2
(sRg)2I(s)/I(0)
Human dystrophin central domain R8-15 fragment Kratky plot 1.104 0 √3 sRg
p(r)
Human dystrophin central domain R8-15 fragment pair distance distribution function Rg: 9.9 nm 0 Dmax: 36 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Human dystrophin central domain R8-15 fragment CUSTOM IN-HOUSE model
Synchrotron SAXS data from solutions of the R8-15 human dystrophin fragment in NaP 10 mM, NaCl 500 mM, EDTA 1 mM, glycerol 2%, pH 7.5 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the SWING beam line at SOLEIL (Saint-Aubin, France) using a CCD AVIEX PCCD170170 detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.1033 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

SEC-SAXS was performed at 15°C using the following parameters: Column: BioSEC5-500Å (4.6 mm id * 300 mm); Flow rate: 0.2 mL/min; Sample injection concentration: 4 mg/mL; Injection volume: 60μL. The data were collected through the SEC peak of the protein as a series of 38 x 1.5 second exposures. The experimental molecular weight was determined from the volume of correlation, Vc.

Human dystrophin central domain R8-15 fragment
Mol. type   Protein
Olig. state   Monomer
Mon. MW   100.2 kDa
 
UniProt   P11532 (1156-1973)
Sequence   FASTA