Human aromatic amino acid decarboxylase is an asymmetric and flexible enzyme: implication in AADC deficiency

Bisello G, Ribeiro R, Perduca M, Belviso B, Polverino de' Laureto P, Giorgetti A, Caliandro R, Bertoldi M, Protein Science (2023) DOI

SASDRA9 – Aromatic-L-amino-acid decarboxylase (AADC) - dopa methylester DME-bound holo form

Aromatic-L-amino-acid decarboxylase (M17V)
MWexperimental 98 kDa
MWexpected 108 kDa
VPorod 154 nm3
log I(s) 2.67×10-1 2.67×10-2 2.67×10-3 2.67×10-4
Aromatic-L-amino-acid decarboxylase (M17V) small angle scattering data  s, nm-1
ln I(s)
Aromatic-L-amino-acid decarboxylase (M17V) Guinier plot ln 2.67×10-1 Rg: 3 nm 0 (3 nm)-2 s2
(sRg)2I(s)/I(0)
Aromatic-L-amino-acid decarboxylase (M17V) Kratky plot 1.104 0 √3 sRg
p(r)
Aromatic-L-amino-acid decarboxylase (M17V) pair distance distribution function Rg: 3.1 nm 0 Dmax: 14.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Aromatic-L-amino-acid decarboxylase (M17V) DAMMIN model
log I(s)
 s, nm-1
Aromatic-L-amino-acid decarboxylase (M17V) OTHER model
Synchrotron SAXS data from solutions of aromatic-L-amino-acid decarboxylase (holo form bound to DME) in 50 mM HEPES, pH 7.4 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.095 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 50.00 μl sample at 4 mg/ml was injected at a 0.33 ml/min flow rate onto a Shodex KW400 series column at 20°C. 600 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.

Aromatic-L-amino-acid decarboxylase (M17V) (AADC)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   53.9 kDa
 
UniProt   P20711
Sequence   FASTA