Inhibition of the Staphylococcus aureus c-di-AMP cyclase DacA by direct interaction with the phosphoglucosamine mutase GlmM.

Tosi T, Hoshiga F, Millership C, Singh R, Eldrid C, Patin D, Mengin-Lecreulx D, Thalassinos K, Freemont P, Gründling A, PLoS Pathog 15(1):e1007537 (2019) Europe PMC

SASDE78 – Solution structure of the diadenylate cyclase/phosphoglucosamine mutase (DacA/GlmM) complex from Staphylococcus aureus

Diadenylate cyclase
Phosphoglucosamine mutase
MWexperimental 131 kDa
MWexpected 138 kDa
VPorod 204 nm3
log I(s) 1.00×10-1 1.00×10-2 1.00×10-3 1.00×10-4
Diadenylate cyclase Phosphoglucosamine mutase small angle scattering data  s, nm-1
ln I(s)
Diadenylate cyclase Phosphoglucosamine mutase Guinier plot ln 1×10-1 Rg: 3.9 nm 0 (3.9 nm)-2 s2
Diadenylate cyclase Phosphoglucosamine mutase Kratky plot 1.104 0 3 sRg
Diadenylate cyclase Phosphoglucosamine mutase pair distance distribution function Rg: 3.9 nm 0 Dmax: 12.1 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Diadenylate cyclase Phosphoglucosamine mutase DAMFILT model

Synchrotron SAXS data from solutions of the DacA/GlmM complex in 30 mM Tris, 150 mM NaCl, pH 7.5 were collected using size-exclusion chromatography SAXS (SEC-SAXS) on the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-to-detector distance of 4.014 m and X-ray wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). SEC-SAXS was performed at 20°C using the following parameters: Column: Superdex 200 5/150 (GE Healthcare); Flow rate: 0.075 mL/min; Sample injection concentration: 12.0 mg/mL; Injection volume: 45 μL. The data obtained through the sample elution peak (collected as consecutive 3 s exposures) were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted and the individual subtracted data sets were scaled and averaged to generate the scattering profile displayed in this entry.

Diadenylate cyclase (DacA)
Mol. type   Protein
Organism   Staphylococcus aureus
Olig. state   Dimer
Mon. MW   19.4 kDa
UniProt   A0A0H2XIU4 (100-269)
Sequence   FASTA
Phosphoglucosamine mutase (GlmM)
Mol. type   Protein
Organism   Staphylococcus aureus
Olig. state   Dimer
Mon. MW   49.7 kDa
UniProt   Q2FEX1
Sequence   FASTA