Modulation of Human Phenylalanine Hydroxylase by 3-Hydroxyquinolin-2(1H)-One Derivatives

Lopes R, Tomé C, Russo R, Paterna R, Leandro J, Candeias N, Gonçalves L, Teixeira M, Sousa P, Guedes R, Vicente J, Gois P, Leandro P, Biomolecules 11(3):462 (2021) DOI

SASDFC7 – Human phenylalanine hydroxylase (hPAH) + 1 mM L-Phe

Phenylalanine-4-hydroxylase
MWexperimental 224 kDa
MWexpected 223 kDa
VPorod 367 nm3
log I(s) 1.50×10-2 1.50×10-3 1.50×10-4 1.50×10-5
Phenylalanine-4-hydroxylase small angle scattering data  s, nm-1
ln I(s)
Phenylalanine-4-hydroxylase Guinier plot ln 1.50×10-2 Rg: 4.0 nm 0 (4.0 nm)-2 s2
(sRg)2I(s)/I(0)
Phenylalanine-4-hydroxylase Kratky plot 1.104 0 √3 sRg
p(r)
Phenylalanine-4-hydroxylase pair distance distribution function Rg: 4.0 nm 0 Dmax: 12.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Phenylalanine-4-hydroxylase DAMMIN model
Phenylalanine-4-hydroxylase DAMFILT model
log I(s)
 s, nm-1
Phenylalanine-4-hydroxylase CORAL model
Synchrotron SAXS data from solutions of Human phenylalanine hydroxylase (hPAH) + 1 mM L-Phe in 0.02 mM Hepes, 0.2 M NaCl,, pH 7 were collected on the B21 beam line at the Diamond Light Source storage ring (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 4 m and at a wavelength of λ = 0.1 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 45.00 μl sample at 12 mg/ml was injected at a 0.16 ml/min flow rate onto a Shodex KW400 series column at 20°C. 541 successive 3 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.

Phenylalanine-4-hydroxylase (PAH)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   55.8 kDa
 
UniProt   P00439 (1-452)
Sequence   FASTA