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

SASDJR8 – Dimeric human ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358

Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358
MWexperimental 76 kDa
MWexpected 68 kDa
VPorod 111 nm3
log I(s) 5.75×103 5.75×102 5.75×101 5.75×100
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358 small angle scattering data  s, nm-1
ln I(s)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358 Guinier plot ln 5.76×103 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358 Kratky plot 1.104 0 √3 sRg
p(r)
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358 pair distance distribution function Rg: 31.2 nm 0 Dmax: 101.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Ganglioside-induced differentiation-associated protein 1, construct GDAP1∆295-358 GASBOR model
Synchrotron SAXS data from solutions of dimeric human GDAP1∆295-358 in 25 mM HEPES, 300 mM NaCl, pH 7.5 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 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 0.50 ml/min flow rate onto a GE Superdex 75 Increase 10/300 column at 15°C. 3000 successive 0.045 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∆295-358 (GDAP1∆295-358)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   34.2 kDa
 
UniProt   Q8TB36 (1-294)
Sequence   FASTA