Dimerization of kringle 1 domain from hepatocyte growth factor/scatter factor provides a potent MET receptor agonist

de Nola G, Leclercq B, Mougel A, Taront S, Simonneau C, Forneris F, Adriaenssens E, Drobecq H, Iamele L, Dubuquoy L, Melnyk O, Gherardi E, de Jonge H, Vicogne J, Life Science Alliance 5(12):e202201424 (2022) DOI

SASDLP9 – Hepatocyte growth factor HGF/SF receptor - fragment MET567 in complex with the minimal hepatocyte growth factor mimic K1K1

minimal hepatocyte growth factor mimic K1K1
Hepatocyte growth factor receptor
MWI(0) 79 kDa
MWexpected 82 kDa
VPorod 136 nm3
log I(s) 7.00×101 7.00×100 7.00×10-1 7.00×10-2
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor small angle scattering data  s, nm-1
ln I(s)
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor Guinier plot ln 7.00×101 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor Kratky plot 1.104 0 √3 sRg
p(r)
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor pair distance distribution function Rg: 3.9 nm 0 Dmax: 14.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor CORAL model
log I(s)
 s, nm-1
minimal hepatocyte growth factor mimic K1K1 Hepatocyte growth factor receptor GASBOR model
Synchrotron SAXS data from solutions of the complex between the HGF/SF receptor - fragment MET567 and the minimal hepatocyte growth factor mimic K1K1 in 25 mM Tris, 150 mM NaCl, pH 7.4 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.0992 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 35.00 μl sample at 12.4 mg/ml was injected at a 0.08 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 20°C. 112 successive 1 second frames were collected through the SEC-elution peak. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

SEC-SAXS experiments were carried out using Nexera High Pressure Liquid/Chromatography (HPLC; Shimadzu) system connected online to SAXS sample capillary. For these experiments, frames corresponding to protein peaks were identified, blank subtracted and averaged using CHROMIXS. Radii of gyration (Rg), molar mass estimates and distance distribution functions P(r) were computed using the ATSAS package in PRIMUS.

minimal hepatocyte growth factor mimic K1K1 (K1K1)
Mol. type   Protein
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   19.2 kDa
Sequence   FASTA
 
Hepatocyte growth factor receptor (MET567)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   62.5 kDa
 
UniProt   P08581 (25-567)
Sequence   FASTA
 
PDB ID   4K3J