The missing part: the Archaeoglobus fulgidus Argonaute forms a functional heterodimer with an N-L1-L2 domain protein.

Manakova E Golovinas E, Pocevičiūtė R, Sasnauskas G, Silanskas A, Rutkauskas D, Jankunec M, Zagorskaitė E, Jurgelaitis E, Grybauskas A, Venclovas Č, Zaremba M, Nucleic Acids Res (2024) Europe PMC

SASDR29 – Single-chain Archaeoglobus fulgidus argonaute fAfAgo-fusion variant in high salt buffer

single-chain full Archaeoglobus fulgidus Argonaute variant
MWI(0) 60 kDa
MWexpected 78 kDa
VPorod 142 nm3
log I(s) 3.72×10-2 3.72×10-3 3.72×10-4 3.72×10-5
single-chain full Archaeoglobus fulgidus Argonaute variant small angle scattering data  s, nm-1
ln I(s)
single-chain full Archaeoglobus fulgidus Argonaute variant Guinier plot ln 3.72×10-2 Rg: 3.1 nm 0 (3.1 nm)-2 s2
(sRg)2I(s)/I(0)
single-chain full Archaeoglobus fulgidus Argonaute variant Kratky plot 1.104 0 √3 sRg
p(r)
single-chain full Archaeoglobus fulgidus Argonaute variant pair distance distribution function Rg: 3.1 nm 0 Dmax: 9.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
single-chain full Archaeoglobus fulgidus Argonaute variant OTHER model
log I(s)
 s, nm-1
single-chain full Archaeoglobus fulgidus Argonaute variant DAMMIN model
log I(s)
 s, nm-1
single-chain full Archaeoglobus fulgidus Argonaute variant DAMMIN model
Synchrotron SAXS data from solutions of a fAfAgo fusion protein in 20 mM TrisHCl pH 7.5, 500 mM NaCl, 5 mM MgCl2, 1 mM DTT, were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 2M 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). One solute concentration of 2.90 mg/ml was measured at 20°C. 20 successive 0.045 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.

The sample was transferred into high salt buffer using an NAP Illustra column and concentrated to 2.9 mg/ml.

single-chain full Archaeoglobus fulgidus Argonaute variant (scfAgo)
Mol. type   Protein
Organism   Archaeoglobus fulgidus
Olig. state   Monomer
Mon. MW   77.9 kDa
Sequence   FASTA