Low-resolution structures of modular nanotransporters shed light on their functional activity

Yury Ryzhykau.

SASDJZ7 – A modular diphtheria toxin/flavohemoprotein nanotransporter (MNT) fused to an epidermal growth factor ligand module (MNT-EGF)

Modular nanotransporter with an epidermal growth factor ligand module
MWexperimental 100 kDa
MWexpected 76 kDa
VPorod 161 nm3
log I(s) 1.90×10-2 1.90×10-3 1.90×10-4 1.90×10-5
Modular nanotransporter with an epidermal growth factor ligand module small angle scattering data  s, nm-1
ln I(s)
Modular nanotransporter with an epidermal growth factor ligand module Guinier plot ln 1.90×10-2 Rg: 4.2 nm 0 (4.2 nm)-2 s2
(sRg)2I(s)/I(0)
Modular nanotransporter with an epidermal growth factor ligand module Kratky plot 1.104 0 3 sRg
p(r)
Modular nanotransporter with an epidermal growth factor ligand module pair distance distribution function Rg: 4.3 nm 0 Dmax: 16 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Modular nanotransporter with an epidermal growth factor ligand module GASBOR model

Synchrotron SAXS data from solutions of MNT-EGF in 150 mM NaCl, 10 mM phosphate buffer saline, pH 8 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 = 0.09918 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 500.00 μl sample at 4.2 mg/ml was injected at a 0.75 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 101 successive 1 second frames were collected through the SEC elution peak of the sample. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The first part of the amino acid sequence of MNT-EGF encompasses sequences homologous to Diphtheria toxin (https://www.uniprot.org/uniprot/P00587) followed by Flavohemoprotein (https://www.uniprot.org/uniprot/P24232). The C-terminal region has a sequence homology to Pro-epidermal growth factor (https://www.uniprot.org/uniprot/P01133).

Modular nanotransporter with an epidermal growth factor ligand module (MNT-EGF)
Mol. type   Protein
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   76.3 kDa
Sequence   FASTA