Identification of a novel tetrameric structure for human apolipoprotein-D.

Kielkopf CS, Low JKK, Mok YF, Bhatia S, Palasovski T, Oakley AJ, Whitten AE, Garner B, Brown SHJ, J Struct Biol 203(3):205-218 (2018) Europe PMC

SASDD83 – Apolipoprotein D (ApoD) tetramer

Apolipoprotein D
MWI(0) 97 kDa
MWexpected 77 kDa
VPorod 169 nm3
log I(s) 3.70×10-2 3.70×10-3 3.70×10-4 3.70×10-5
Apolipoprotein D small angle scattering data  s, nm-1
ln I(s)
Apolipoprotein D Guinier plot ln 3.70×10-2 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
Apolipoprotein D Kratky plot 1.104 0 √3 sRg
p(r)
Apolipoprotein D pair distance distribution function Rg: 3.4 nm 0 Dmax: 11.0 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Apolipoprotein D SASREF CV model
log I(s)
 s, nm-1
Apolipoprotein D SASREF CV model
Synchrotron SAXS data from solutions of apolipoprotein D (ApoD) tetramer in 50 mM Na Phosphate, 150 mM NaCl, 3% glycerol, pH 7.4 were collected on the SAXS/WAXS beam line at the Australian Synchrotron (Melbourne, Australia) using a Pilatus 1M detector at a wavelength of λ = 0.10322 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). The SAXS data were collected using size-exclusion chromatography SAXS (SEC-SAXS).

Additional SEC-SAXS information: SEC-column: GE Healthcare Superdex S200 5/150; Loading concentration (measured using BCA assay): 7.5 mg/ml; Injection volume : 50 µl; Flow rate: 0.45 ml/min; Number of frames averaged over the elution peak: 15. The starting pdb model was the apo-form of apoD with modelled glycans (Oakley et al. 2012). The refined SASREF CV model incorporated contact restraints derived from crosslinking mass spectrometry (crosslinker BS3) that are included in the full entry zip archive (32 Ang between: a, K55-K55; b, K156-K156; c, K155-K156 and; d, K144-K156.)

Apolipoprotein D (ApoD)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Tetramer
Mon. MW   19.3 kDa
 
UniProt   P05090 (21-189)
Sequence   FASTA