Self-Assembling Supramolecular Nanostructures Constructed from de Novo Extender Protein Nanobuilding Blocks.

Kobayashi N, Inano K, Sasahara K, Sato T, Miyazawa K, Fukuma T, Hecht MH, Song C, Murata K, Arai R, ACS Synth Biol 7(5):1381-1394 (2018) PubMed

SASDD46 – Artificially designed de novo protein esPN-Block (HL4) heterocomplex, e1s2 (HL4)

extender PN-Block (HL4)
stopper PN-Block (WA20)
MWI(0) 58 kDa
MWexpected 53 kDa
log I(s) 3.50×10-2 3.50×10-3 3.50×10-4 3.50×10-5
extender PN-Block (HL4) stopper PN-Block (WA20) small angle scattering data  s, nm-1
ln I(s)
extender PN-Block (HL4) stopper PN-Block (WA20) Guinier plot ln 3.50×10-2 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
extender PN-Block (HL4) stopper PN-Block (WA20) Kratky plot 1.104 0 3 sRg
p(r)
extender PN-Block (HL4) stopper PN-Block (WA20) pair distance distribution function Rg: 4.1 nm 0 Dmax: 15 nm

Data validation


Fits and models


log I(s)
 s, nm-1
extender PN-Block (HL4) stopper PN-Block (WA20) DAMMIN model

log I(s)
 s, nm-1
extender PN-Block (HL4) stopper PN-Block (WA20) PYMOL model

Synchrotron SAXS data from solutions of an artificially designed de novo protein, esPN-Block (HL4) heterocomplex e1s2 (HL4), in 20 mM HEPES, 100 mM NaCl, 200 mM ArgHCl, 10% v/v glycerol, pH 7.5 were collected on the BL-10C beam line at the Photon Factory (PF), High Energy Acceleration Research Organization (KEK; Tsukuba, Japan) using a Pilatus 2M detector at a sample-detector distance of 1.1 m and at a wavelength of λ = 0.1488 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.60 mg/ml was measured at 20°C. 30 successive 5 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

extender PN-Block (HL4) (ePN-Block (HL4))
Mol. type   Protein
Organism   artificially designed de novo protein
Olig. state   Monomer
Mon. MW   27.5 kDa
Sequence   FASTA
 
stopper PN-Block (WA20) (sPN-Block (WA20))
Mol. type   Protein
Organism   artificially designed de novo protein
Olig. state   Dimer
Mon. MW   12.5 kDa
Sequence   FASTA
 
PDB code   3VJF