Structural basis of CDNF interaction with the UPR regulator GRP78.

Graewert MA, Volkova M, Jonasson K, Määttä JAE, Gräwert T, Mamidi S, Kulesskaya N, Evenäs J, Johnsson RE, Svergun D, Bhattacharjee A, Huttunen HJ, Nat Commun 15(1):8175 (2024) Europe PMC

SASDNW9 – Glucose-regulated protein 78, nucleotide-binding domain

Endoplasmic reticulum chaperone BiP
MWexperimental 45 kDa
MWexpected 42 kDa
VPorod 70 nm3
log I(s) 1.97×103 1.97×102 1.97×101 1.97×100
Endoplasmic reticulum chaperone BiP small angle scattering data  s, nm-1
ln I(s)
Endoplasmic reticulum chaperone BiP Guinier plot ln 1.97×103 Rg: 2.2 nm 0 (2.2 nm)-2 s2
(sRg)2I(s)/I(0)
Endoplasmic reticulum chaperone BiP Kratky plot 1.104 0 √3 sRg
p(r)
Endoplasmic reticulum chaperone BiP pair distance distribution function Rg: 2.2 nm 0 Dmax: 6.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Endoplasmic reticulum chaperone BiP PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of glucose-regulated protein 78, nucleotide-binding domain in phosphate buffered saline, pH 7.2 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 10.00 μl sample at 10 mg/ml was injected at a 0.35 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 30 successive 0.500 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.

Endoplasmic reticulum chaperone BiP (GRP78-NBD)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   42.0 kDa
 
UniProt   P11021 (26-405)
Sequence   FASTA
 
PDB ID   3IUC