Structural basis for the inhibition of the Bacillus subtilis c-di-AMP cyclase CdaA by the phosphoglucomutase GlmM

Pathania M, Tosi T, Millership C, Hoshiga F, Morgan R, Freemont P, Gründling A, Journal of Biological Chemistry :101317 (2021) DOI

SASDMQ5 – Bacillus subtilis complex of diadenylate cyclase (CdaA:cytoplasmic domain) in complex with a truncated version of phosphoglucosamine mutase (GlmMF369)

Cyclic di-AMP synthase CdaA
Phosphoglucosamine mutase
MWexperimental 98 kDa
MWexpected 127 kDa
VPorod 156 nm3
log I(s) 1.20×10-1 1.20×10-2 1.20×10-3 1.20×10-4
Cyclic di-AMP synthase CdaA Phosphoglucosamine mutase small angle scattering data  s, nm-1
ln I(s)
Cyclic di-AMP synthase CdaA Phosphoglucosamine mutase Guinier plot ln 1.20×10-1 Rg: 3.7 nm 0 (3.7 nm)-2 s2
(sRg)2I(s)/I(0)
Cyclic di-AMP synthase CdaA Phosphoglucosamine mutase Kratky plot 1.104 0 √3 sRg
p(r)
Cyclic di-AMP synthase CdaA Phosphoglucosamine mutase pair distance distribution function Rg: 3.8 nm 0 Dmax: 15.1 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of diadenylate cyclase (CdaA:cytoplasmic domain) in complex with a truncated version of phosphoglucosamine mutase (GlmMF369) in 30 mM Tris, 150 mM NaCl, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a wavelength of λ = 0.1 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 45.00 μl sample at 10 mg/ml was injected at a 0.16 ml/min flow rate onto a Shodex KW403 column at 20°C. 620 successive 3 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.

Sample detector distance = UNKNOWN

Cyclic di-AMP synthase CdaA (Diadenylate cyclase)
Mol. type   Protein
Organism   Bacillus subtilis (strain 168)
Olig. state   Dimer
Mon. MW   22.0 kDa
 
UniProt   Q45589 (97-273)
Sequence   FASTA
 
Phosphoglucosamine mutase (GlmMF369)
Mol. type   Protein
Organism   Bacillus subtilis
Olig. state   Dimer
Mon. MW   41.3 kDa
 
UniProt   O34824 (1-369)
Sequence   FASTA