Determinants of receptor tyrosine phosphatase homophilic adhesion: structural comparison of PTPRK and PTPRM extracellular domains

Hay I, Shamin M, Caroe E, Mohammed A, Svergun D, Jeffries C, Graham S, Sharpe H, Deane J, Journal of Biological Chemistry :102750 (2022) DOI

SASDPF3 – Receptor-type tyrosine-protein phosphatase kappa extracellular domains (PTPRK-ECDs)

Receptor-type tyrosine-protein phosphatase kappa
MWexperimental 113 kDa
MWexpected 82 kDa
VPorod 252 nm3
log I(s) 5.23×10-2 5.23×10-3 5.23×10-4 5.23×10-5
Receptor-type tyrosine-protein phosphatase kappa small angle scattering data  s, nm-1
ln I(s)
Receptor-type tyrosine-protein phosphatase kappa Guinier plot ln 5.23×10-2 Rg: 7.0 nm 0 (7.0 nm)-2 s2
(sRg)2I(s)/I(0)
Receptor-type tyrosine-protein phosphatase kappa Kratky plot 1.104 0 3 sRg
p(r)
Receptor-type tyrosine-protein phosphatase kappa pair distance distribution function Rg: 7.5 nm 0 Dmax: 26 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Receptor-type tyrosine-protein phosphatase kappa DAMMIN model
Receptor-type tyrosine-protein phosphatase kappa DAMMIN model

log I(s)
 s, nm-1
Receptor-type tyrosine-protein phosphatase kappa CORAL model

Synchrotron SAXS data from solutions of the receptor-type tyrosine-protein phosphatase kappa extracellular domains (PTPRK-ECDs) in 50 mM MES, 250 mM NaCl, 3% v/v glycerol, pH 6 were collected on the EMBL P12 beam line at PETRA III (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 40.00 μl sample at 2.3 mg/ml was injected at a 0.35 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 2880 successive 0.250 second frames were collected through the entire column elution; 47 data frames from the main sample peak were used for averaging. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted (processed using CHROMIXS).

The quoted experimental molecular weight of 113 kDa was obtained using a Bayesian inference concentration independent method from the SAXS data. The corresponding experimental MALLS molecular weight estimate is 105 kDa, that includes a conjugate analysis of the protein molecular weight of (79 kDa) and glycan component (26 kDa). The corresponding SEC-SAXS and SEC-MALLS-QELS data as well as the results from DAMMIN ab initio, and hybrid CORAL/Alphafold/Xray crystal structure rigid-body modelling are made available in the full entry zip archive.

Receptor-type tyrosine-protein phosphatase kappa (PTPRK)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   81.9 kDa
 
UniProt   Q15262 (28-752)
Sequence   FASTA