Structural Insights into the Intracellular Region of the Human Magnesium Transport Mediator CNNM4.

Giménez-Mascarell P, Oyenarte I, González-Recio I, Fernández-Rodríguez C, Corral-Rodríguez MÁ, Campos-Zarraga I, Simón J, Kostantin E, Hardy S, Díaz Quintana A, Zubillaga Lizeaga M, Merino N, Diercks T, Blanco FJ, Díaz Moreno I, Martínez-Chantar ML, Tremblay ML, Müller D, Siliqi D, Martínez-Cruz LA, Int J Mol Sci 20(24) (2019) Europe PMC

SASDES8 – Intracellular region of the human Cyclin and CBS domain divalent metal cation transport mediator-4 in complex with MgATP

CNNM4_BAT-cNMP-Ctail
MWexperimental 94 kDa
MWexpected 96 kDa
VPorod 149 nm3
log I(s) 4.18×10-2 4.18×10-3 4.18×10-4 4.18×10-5
CNNM4_BAT-cNMP-Ctail small angle scattering data  s, nm-1
ln I(s)
CNNM4_BAT-cNMP-Ctail Guinier plot ln 4.19×10-2 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
CNNM4_BAT-cNMP-Ctail Kratky plot 1.104 0 3 sRg
p(r)
CNNM4_BAT-cNMP-Ctail pair distance distribution function Rg: 3.9 nm 0 Dmax: 14.3 nm

Data validation


Fits and models


log I(s)
 s, nm-1
CNNM4_BAT-cNMP-Ctail CORAL model

log I(s)
 s, nm-1
CNNM4_BAT-cNMP-Ctail DAMFILT model

Synchrotron SAXS data from solutions of Intracellular region of the human Cyclin and CBS domain divalent metal cation transport mediator-4 in complex with MgATP in HEPES buffer pH 7.4, 200 mM NaCl, 1mM DTT, pH 7.4 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the B21 camera at the Diamond Light Source storage ring (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). The data obtained through the sample elution peak (collected as 620 successive 3 second frames) were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted and the individual subtracted data sets were scaled and averaged to generate the scattering profile displayed in this entry.

SEC-SAXS was performed at 20°C using the following parameters: Column: Shodex KW-403 ; Flow rate: 0.08 mL/min; Total acquisition time: 1860s; Sample injection concentration: 4.7 mg/mL; Injection volume: 45μL.

CNNM4_BAT-cNMP-Ctail (CNNM4_intra)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   47.9 kDa
 
UniProt   Q6P4Q7
Sequence   FASTA