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) Europe PMC

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.

Storage temperature = UNKNOWN

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