Structure of the Complete Dimeric Human GDAP1 Core Domain Provides Insights into Ligand Binding and Clustering of Disease Mutations

Nguyen G, Sutinen A, Raasakka A, Muruganandam G, Loris R, Kursula P, Frontiers in Molecular Biosciences 7 (2021) DOI

SASDJW8 – Monomeric human ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A

Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A
MWexperimental 35 kDa
MWexpected 35 kDa
VPorod 59 nm3
log I(s) 1.03×10-1 1.03×10-2 1.03×10-3 1.03×10-4
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A small angle scattering data  s, nm-1
ln I(s)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A Guinier plot ln 1.03×10-1 Rg: 2.5 nm 0 (2.5 nm)-2 s2
(sRg)2I(s)/I(0)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A Kratky plot 1.104 0 √3 sRg
p(r)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A pair distance distribution function Rg: 2.5 nm 0 Dmax: 8.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A GASBOR model
Synchrotron SAXS data from solutions of monomeric human GDAP1∆303-358 (Y29E/C88A) 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 15°C. 480 successive 0.990 second frames were collected through the SEC elution profile. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted from the sample elution peak frames to generate the final SAXS profile.

Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆303-358, mutant Y29E/C88A (GDAP1∆303-358 mutant)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   35.1 kDa
 
UniProt   Q8TB36 (1-302)
Sequence   FASTA