Structural and Biochemical Characterization of Aldehyde Dehydrogenase 12, the Last Enzyme of Proline Catabolism in Plants.

Korasick DA, Končitíková R, Kopečná M, Hájková E, Vigouroux A, Moréra S, Becker DF, Šebela M, Tanner JJ, Kopečný D, J Mol Biol (2018) Europe PMC

SASDE96 – Aldehyde dehydrogenase 12 from Zea mays Extrapolated to Infinite Dilution

Aldehyde dehydrogenase 12
MWexperimental 250 kDa
MWexpected 242 kDa
VPorod 351 nm3
log I(s) 9.26×101 9.26×100 9.26×10-1 9.26×10-2
Aldehyde dehydrogenase 12 small angle scattering data  s, nm-1
ln I(s)
Aldehyde dehydrogenase 12 Guinier plot ln 9.26×101 Rg: 4.1 nm 0 (4.1 nm)-2 s2
Aldehyde dehydrogenase 12 Kratky plot 1.104 0 3 sRg
Aldehyde dehydrogenase 12 pair distance distribution function Rg: 4.0 nm 0 Dmax: 14.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Aldehyde dehydrogenase 12 ALLOSMOD model

log I(s)
 s, nm-1
Aldehyde dehydrogenase 12 ALLOSMOD model

Synchrotron SAXS data from solutions of aldehyde dehydrogenase in 50 mM Tris-HCl, 50 mM NaCl, 0.5 mM TCEP, and 5% (v/v) glycerol, pH 7.8 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS, Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.127 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). 33 successive 0.300 second frames were collected at a sample temperature of 10°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 concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.

This is the background subtracted experimental SAXS data extrapolated to infinite dilution. The original experiment consisted of static SAXS samples collected at 3 protein concentrations in the range of 0.9 - 2.6 mg/mL. The data were extrapolated to infinite dilution to account for sample aggregation in the two higher concentration samples. The corresponding model fits of the tetramer (top) and dimer (bottom) demonstrate that the dimer does not fit the SAXS data.

Aldehyde dehydrogenase 12
Mol. type   Protein
Organism   Zea mays
Olig. state   Tetramer
Mon. MW   60.4 kDa
UniProt   A0A2H4PMI3 (17-549)
Sequence   FASTA