Antiviral signalling by a cyclic nucleotide activated CRISPR protease.

Rouillon C, Schneberger N, Chi H, Blumenstock K, Da Vela S, Ackermann K, Moecking J, Peter MF, Boenigk W, Seifert R, Bode BE, Schmid-Burgk JL, Svergun D, Geyer M, White MF, Hagelueken G, Nature 614(7946):168-174 (2023) Europe PMC

SASDQN4 – Complex of CRISPR associated Lon protease (CalpL) with a 10 kDa C-terminal fragment of CalpT a putative MazF-like toxin (CalpT10)

SAVED domain-containing protein
Uncharacterized protein (putative MazF-like toxin)
MWexperimental 62 kDa
MWexpected 66 kDa
VPorod 93 nm3
log I(s) 7.37×102 7.37×101 7.37×100 7.37×10-1
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) small angle scattering data  s, nm-1
ln I(s)
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) Guinier plot ln 7.38×102 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) Kratky plot 1.104 0 √3 sRg
p(r)
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) pair distance distribution function Rg: 3.5 nm 0 Dmax: 11.7 nm

Data validation

Fits and models

log I(s)
 s, nm-1
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) CUSTOM IN-HOUSE model
log I(s)
 s, nm-1
SAVED domain-containing protein Uncharacterized protein (putative MazF-like toxin) MONSA model
Synchrotron SAXS data from solutions of CRISPR associated Lon protease (CalpL) bound to a 10 kDa C-terminal fragment of CalpT in 20 mM Tris, 50 mM NaCl, pH 8 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.124 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 40.00 μl sample at 7.6 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 900 successive 1 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.

SAVED domain-containing protein (CalpL)
Mol. type   Protein
Organism   Sulfurihydrogenibium sp. (strain YO3AOP1)
Olig. state   Monomer
Mon. MW   57.7 kDa
 
UniProt   B2V8L9 (2-496)
Sequence   FASTA
 
Uncharacterized protein (putative MazF-like toxin) (CalpT10)
Mol. type   Protein
Organism   Sulfurihydrogenibium sp. (strain YO3AOP1)
Olig. state   Monomer
Mon. MW   8.7 kDa
 
UniProt   B2V8L8 (196-271)
Sequence   FASTA