Structural characterization of the third scavenger receptor cysteine-rich domain of murine Neurotrypsin

Canciani A, Catucci G, Forneris F, Protein Science (2019) DOI

SASDES5 – Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 (mmNT-SRCR3) - from SEC-SAXS

Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3
MWI(0) 12 kDa
MWexpected 13 kDa
VPorod 24 nm3
log I(s) 1.34×101 1.34×100 1.34×10-1 1.34×10-2
Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 small angle scattering data  s, nm-1
ln I(s)
Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 Guinier plot ln 1.35×101 Rg: 1.5 nm 0 (1.5 nm)-2 s2
(sRg)2I(s)/I(0)
Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 Kratky plot 1.104 0 3 sRg
p(r)
Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 pair distance distribution function Rg: 1.5 nm 0 Dmax: 4.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 PDB model

Synchrotron SAXS data, I(s) vs s (where s = 4πsinθ/λ, 2θ is the scattering angle and λ the X-ray wavelength), were collected from a sample of mmNT-SRCR3 using continuous-flow size-exclusion chromatography SAXS (SEC-SAXS) on the BM29 beam line of the ESRF (Grenoble France) coupled to a Nexera High Pressure Liquid/Chromatography (HPLC; Shimadzu) system. Data were collected using a Pilatus 1M detector at a sample-detector distance of 2.87 m (s-range; 0.03–4.5 nm-1) at a wavelength of λ = 0.09919 nm (12.5 keV). The SEC mobile phase consisted of 25 mM HEPES/NaOH, 0.1 M NaCl, pH 8, (20°C). The protein was concentrated to 15 mg/mL and a 15 µL of sample was run on a Superdex 75 Increase 3.2/300 column (GE Healthcare). Analysis and of the scattering intensities measured from the SEC-elution sample peak (22 successive 1 s frames) and the subtraction of intensities measured from an appropriate solvent-blank were was performed using CHROMIXS of the ATSAS suite. Comparison of in-solution scattering to crystallographic data was carried out with CRYSOL.

Mouse Neurotrypsin Scavenger Receptor Cysteine-Rich Domain 3 (mmNT-SRCR3)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   13.2 kDa
 
UniProt   O08762 (383-494)
Sequence   FASTA
 
PDB code   6H8M