Critical Structural Defects Explain Filamin A Mutations Causing Mitral Valve Dysplasia

Haataja T, Capoulade R, Lecointe S, Hellman M, Merot J, Permi P, Pentikäinen U, Biophysical Journal (2019) DOI

SASDFD3 – Filamin A Ig-like domains 4-6 (FLNa4-6)

Filamin A Ig-like domains 4-6
MWexperimental 23 kDa
MWexpected 32 kDa
VPorod 41 nm3
log I(s) 2.37×101 2.37×100 2.37×10-1 2.37×10-2
Filamin A Ig-like domains 4-6 small angle scattering data  s, nm-1
ln I(s)
Filamin A Ig-like domains 4-6 Guinier plot ln 2.37×101 Rg: 2.7 nm 0 (2.7 nm)-2 s2
(sRg)2I(s)/I(0)
Filamin A Ig-like domains 4-6 Kratky plot 1.104 0 3 sRg
p(r)
Filamin A Ig-like domains 4-6 pair distance distribution function Rg: 2.8 nm 0 Dmax: 9.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Filamin A Ig-like domains 4-6 DAMFILT model

log I(s)
 s, nm-1
Filamin A Ig-like domains 4-6 SASREF model

Synchrotron SAXS data from solutions of Filamin A Ig-like domains 4-6 in 20 mM Tris, 100 mM NaCl, 1 mM DTT, pH 8 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2 and 4 mg/ml were measured at 20°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Wild type three-domain fragment of human Filamin A Ig-like domains 4-6 (FLNa4-6). There are two extra amino acids, Met and Ser, in the very N-terminus of the polypeptide that have resulted from TEV-cleavage of the GST fusion tag during the protein purification.

Filamin A Ig-like domains 4-6 (FLNa4-6)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   31.5 kDa
 
UniProt   P21333 (574-869)
Sequence   FASTA
 
PDB code   4M9P