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