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