Structure of the Human Cation-Independent Mannose 6-Phosphate/IGF2 Receptor Domains 7–11 Uncovers the Mannose 6-Phosphate Binding Site of Domain 9

Bochel A, Williams C, McCoy A, Hoppe H, Winter A, Nicholls R, Harlos K, Jones E, Berger I, Hassan A, Crump M, Structure (2020) DOI

SASDH59 – Human Cation-independent mannose-6-phosphate receptor domains 9-10 wih mannose-6-phosphate

Cation-independent mannose-6-phosphate receptor
MWI(0) 79 kDa
MWexpected 67 kDa
VPorod 88 nm3
log I(s) 9.72×10-2 9.72×10-3 9.72×10-4 9.72×10-5
Cation-independent mannose-6-phosphate receptor small angle scattering data  s, nm-1
ln I(s)
Cation-independent mannose-6-phosphate receptor Guinier plot ln 9.72×10-2 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
Cation-independent mannose-6-phosphate receptor Kratky plot 1.104 0 √3 sRg
p(r)
Cation-independent mannose-6-phosphate receptor pair distance distribution function Rg: 3.2 nm 0 Dmax: 9.1 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Cation-independent mannose-6-phosphate receptor OTHER model
Synchrotron SAXS data from solutions of Human D9-10 protein with a 100 fold excess of mannose-6-phosphate in 25 mM Tris 150 mM NaCl, pH 7.5, were collected using in-line SEC-SAXS at the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In line SEC-SAXS was performed using an Agilent 1200 HPLC system equipped with a 2.4 mL Superdex S200 column (GE Healthcare). A sample concentration of 6.5 mg/ml was loaded onto the column at a flow rate of 0.075 mL/min. Data frames were collected at 25°C for the entire column elution with an exposure time of 3 seconds per frame through an angular s-range of 0.03-3.4 nm-1. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted using data frames measured from the flow-through column buffer with no macromolecular sample present.

Cation-independent mannose-6-phosphate receptor (IGF2R - Domain 9-10)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   33.6 kDa
 
UniProt   P11717 (1222-1510)
Sequence   FASTA