Unfolding of monomeric lipoprotein lipase by ANGPTL4: Insight into the regulation of plasma triglyceride metabolism

Kristensen K, Leth-Espensen K, Mertens H, Birrane G, Meiyappan M, Olivecrona G, Jørgensen T, Young S, Ploug M, Proceedings of the National Academy of Sciences :201920202 (2020) DOI

SASDHF4 – Lipoprotein lipase-GPIHBP1-monoclonal antibody (5D2) complex

Lipoprotein lipase
Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1
Monoclonal Antibody Fragment 5D2
MWI(0) 100 kDa
MWexpected 112 kDa
VPorod 187 nm3
log I(s) 2.30×10-2 2.30×10-3 2.30×10-4 2.30×10-5
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 small angle scattering data  s, nm-1
ln I(s)
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 Guinier plot ln 2.30×10-2 Rg: 4.9 nm 0 (4.9 nm)-2 s2
(sRg)2I(s)/I(0)
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 Kratky plot 1.104 0 3 sRg
p(r)
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 pair distance distribution function Rg: 4.8 nm 0 Dmax: 17.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 CORAL model

log I(s)
 s, nm-1
Lipoprotein lipase Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 Monoclonal Antibody Fragment 5D2 DAMMIN model

Synchrotron SAXS data from solutions of Lipoprotein lipase-GPIHBP1-monoclonal antibody (5D2) complex in 10 mM Tris, 150 mM NaCl, 4 mM CaCL2, 10% (v/v) Glycerol, 0.05% 0.8mM CHAPS, ,0.05 % (v/v) NaN3, 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 m and at a wavelength of λ = 0.124 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 3.4 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 10°C. 900 successive 60 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.

Both LPL and GPIHBP1 components of the ternary complex are glycosylated. Based on Mass Spectrometry data the expected glycosylation composition of the scattering particle are 1 x C40N2O29H67 (GPIHBP1) and 2 x C68N4O49H113 (LPL). Thus the expected molecular mass for the ternary complex is ~ 111 kDa.

Lipoprotein lipase (LPL)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   50.4 kDa
 
UniProt   P05868 (29-475)
Sequence   FASTA
 
Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   14.7 kDa
 
UniProt   Q8IV16 (21-151)
Sequence   FASTA
 
Monoclonal Antibody Fragment 5D2 (Fab-5D2)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   46.9 kDa
Sequence   FASTA