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
(sRg)2I(s)/I(0)
Diadenylate cyclase Phosphoglucosamine mutase Kratky plot 1.104 0 √3 sRg
p(r)
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.

Number of frames = UNKNOWN. SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

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