| MWI(0) | 100 | kDa |
| MWexpected | 112 | kDa |
| VPorod | 187 | nm3 |
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log I(s)
2.30×10-2
2.30×10-3
2.30×10-4
2.30×10-5
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s, nm-1
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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) DOISASDHF4 – Lipoprotein lipase-GPIHBP1-monoclonal antibody (5D2) complex
Lipoprotein lipaseGlycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1
Monoclonal Antibody Fragment 5D2
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Fits and models
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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
(DESY; 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. |
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