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

SASDQS6 – Glucose-regulated protein 78 nucleotide-binding domain (GRP78-NBD) in complex with the C-terminal of Cerebral dopamine neurotrophic factor (C-CDNF)

Endoplasmic reticulum chaperone BiP
C-terminal of the Cerebral dopamine neurotrophic factor
MWexperimental 50 kDa
MWexpected 49 kDa
VPorod 66 nm3
log I(s) 2.47×103 2.47×102 2.47×101 2.47×100
Endoplasmic reticulum chaperone BiP C-terminal of the Cerebral dopamine neurotrophic factor small angle scattering data  s, nm-1
ln I(s)
Endoplasmic reticulum chaperone BiP C-terminal of the Cerebral dopamine neurotrophic factor Guinier plot ln 2.47×103 Rg: 2.4 nm 0 (2.4 nm)-2 s2
(sRg)2I(s)/I(0)
Endoplasmic reticulum chaperone BiP C-terminal of the Cerebral dopamine neurotrophic factor Kratky plot 1.104 0 √3 sRg
p(r)
Endoplasmic reticulum chaperone BiP C-terminal of the Cerebral dopamine neurotrophic factor pair distance distribution function Rg: 2.3 nm 0 Dmax: 6.9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Endoplasmic reticulum chaperone BiP C-terminal of the Cerebral dopamine neurotrophic factor SREFLEX model
Synchrotron SAXS data from solutions of GRP78-NBD in complex with C-CDNF 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 25.00 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. 1800 successive 0.500 second frames were collected from the SEC-elution, where 12 frames were processed through the sample elution peak. 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
 
C-terminal of the Cerebral dopamine neurotrophic factor (C-CDNF)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   7.0 kDa
 
UniProt   Q49AH0 (127-187)
Sequence   FASTA