Structure of ATP citrate lyase and the origin of citrate synthase in the Krebs cycle

Verschueren K, Blanchet C, Felix J, Dansercoer A, De Vos D, Bloch Y, Van Beeumen J, Svergun D, Gutsche I, Savvides S, Verstraete K, Nature (2019) DOI

SASDE56 – Human ATP-citrate synthase (ACLY) in HBS + Citrate + Coenzyme-A

ATP-citrate synthase
MWexperimental 480 kDa
MWexpected 458 kDa
VPorod 709 nm3
log I(s) 1.18×104 1.18×103 1.18×102 1.18×101
ATP-citrate synthase small angle scattering data  s, nm-1
ln I(s)
ATP-citrate synthase Guinier plot ln 1.18×104 Rg: 5.8 nm 0 (5.8 nm)-2 s2
(sRg)2I(s)/I(0)
ATP-citrate synthase Kratky plot 1.104 0 3 sRg
p(r)
ATP-citrate synthase pair distance distribution function Rg: 5.8 nm 0 Dmax: 16.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
ATP-citrate synthase SASREF CV model

log I(s)
 s, nm-1
ATP-citrate synthase MULTIFOXS model

Synchrotron SAXS data from solutions of ATP-citrate synthase (ACLY) in 20mM HEPES, 150mM NaCl, 50mM Tris, 20mM citrate, 2mM coenzyme-A, pH 7.2 were collected on the EMBL P12 beam line at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.124 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Size-exclusion chromatography SAXS (SEC-SAXS) was employed using a sample injection concentration of 24.00 mg/ml. Data were measured at through the SEC elution at 20°C. 900 successive 1 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of an appropriate solvent-blank was subtracted using the CHROMIXS SEC-SAXS analysis package.

ATP-citrate synthase
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   114.6 kDa
 
UniProt   P53396-1
Sequence   FASTA