An Integrative Structural Biology Analysis of Von Willebrand Factor Binding and Processing by ADAMTS-13 in Solution.

Del Amo-Maestro L, Sagar A, Pompach P, Goulas T, Scavenius C, Ferrero DS, Castrillo-Briceño M, Taulés M, Enghild JJ, Bernadó P, Gomis-Rüth FX, J Mol Biol 433(13):166954 (2021) Europe PMC

SASDKV8 – ADAMTS-13 bound to von Willebrand Factor peptide

von Willebrand factor
A disintegrin and metalloproteinase with thrombospondin motifs 13
MWexperimental 114 kDa
MWexpected 82 kDa
VPorod 170 nm3
log I(s) 8.68×103 8.68×102 8.68×101 8.68×100
von Willebrand factor A disintegrin and metalloproteinase with thrombospondin motifs 13 small angle scattering data  s, nm-1
ln I(s)
von Willebrand factor A disintegrin and metalloproteinase with thrombospondin motifs 13 Guinier plot ln 8.69×103 Rg: 4.4 nm 0 (4.4 nm)-2 s2
(sRg)2I(s)/I(0)
von Willebrand factor A disintegrin and metalloproteinase with thrombospondin motifs 13 Kratky plot 1.104 0 √3 sRg
p(r)
von Willebrand factor A disintegrin and metalloproteinase with thrombospondin motifs 13 pair distance distribution function Rg: 4.5 nm 0 Dmax: 16.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
ADAMTS-13 bound to von Willebrand Factor peptide Rg histogram Rg, nm
Synchrotron SAXS data from solutions of ADAMTS-13 bound to von Willebrand Factor peptide in 10 mM HEPES, 150 mM NaCl, pH 7.4 were collected on the EMBL P12 beam line at PETRA III (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 100.00 μl sample at 6 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 5/150 column at 20°C. 600 successive 1 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.

SEC-SAXS data collected on the complex of ADAMTS13 and vWF.

von Willebrand factor (VWF(1596-1668))
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   11.2 kDa
 
UniProt   P04275 (1596-1668)
Sequence   FASTA
 
A disintegrin and metalloproteinase with thrombospondin motifs 13 (ADAMTS13)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   70.4 kDa
 
UniProt   Q76LX8 (75-685)
Sequence   FASTA