Structural insights into Charcot-Marie-Tooth disease-linked mutations in human GDAP1.

Sutinen A, Nguyen GTT, Raasakka A, Muruganandam G, Loris R, Ylikallio E, Tyynismaa H, Bartesaghi L, Ruskamo S, Kursula P, FEBS Open Bio (2022) Europe PMC

SASDND6 – Human ganglioside-induced differentiation-associated protein 1 (GDAP1) missense mutant H123R, construct GDAP1∆302-358

Ganglioside-induced differentiation-associated protein 1 (H123R)
MWexperimental 74 kDa
MWexpected 70 kDa
VPorod 107 nm3
log I(s) 1.18×10-1 1.18×10-2 1.18×10-3 1.18×10-4
Ganglioside-induced differentiation-associated protein 1 (H123R) small angle scattering data  s, nm-1
ln I(s)
Ganglioside-induced differentiation-associated protein 1 (H123R) Guinier plot ln 1.18×10-1 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
Ganglioside-induced differentiation-associated protein 1 (H123R) Kratky plot 1.104 0 3 sRg
p(r)
Ganglioside-induced differentiation-associated protein 1 (H123R) pair distance distribution function Rg: 3.1 nm 0 Dmax: 9.9 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Ganglioside-induced differentiation-associated protein 1 (H123R) GASBOR model

Synchrotron SAXS data from solutions of GDAP1∆302-358 (missense mutant H123R) in 25 mM HEPES, 300 mM NaCl, pH 7.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a Eiger 4M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 10 mg/ml was injected at a 0.20 ml/min flow rate onto a Agilent Bio SEC-3, 300 Å column at 20°C. 779 successive 0.990 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 protein construct used for SAXS represents the cytosolic part of GDAP1 with missense point mutation H123R, lacking mitochondrial outer membrane transmembrane helix. The sample was prepared at Oulu in sample buffer and transported to SWING beam line in dry ice, prior to measurement.

Ganglioside-induced differentiation-associated protein 1 (H123R) (GDAP1 (H123R))
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   35.1 kDa
 
UniProt   Q8TB36 (1-301)
Sequence   FASTA
 
ALPHAFOLD ID   Q8TB36