Thermodynamics, cooperativity and stability of the tetracycline repressor (TetR) upon tetracycline binding.

Palm GJ, Buchholz I, Werten S, Girbardt B, Berndt L, Delcea M, Hinrichs W, Biochim Biophys Acta Proteins Proteom :140404 (2020) Europe PMC

SASDFE5 – Tetracycline repressor (class D), TetR(D), in complex with 5a,6-anhydrotetracycline (ATc)

Tetracycline repressor (class D)
5a,6-anhydrotetracycline
MWI(0) 43 kDa
MWexpected 47 kDa
VPorod 77 nm3
log I(s) 7.68×103 7.68×102 7.68×101 7.68×100
Tetracycline repressor (class D) 5a,6-anhydrotetracycline small angle scattering data  s, nm-1
ln I(s)
Tetracycline repressor (class D) 5a,6-anhydrotetracycline Guinier plot ln 7.68×103 Rg: 2.6 nm 0 (2.6 nm)-2 s2
(sRg)2I(s)/I(0)
Tetracycline repressor (class D) 5a,6-anhydrotetracycline Kratky plot 1.104 0 √3 sRg
p(r)
Tetracycline repressor (class D) 5a,6-anhydrotetracycline pair distance distribution function Rg: 2.4 nm 0 Dmax: 6.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Tetracycline repressor (class D) 5a,6-anhydrotetracycline PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of TetR(D), in complex with ATc in 50 mM Tris/HCl 150 mM NaCl 10 mM MgCl2, pH 8 were collected on the EMBL P12 beam line at PETRA III (Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.124 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 1.4 and 1.4 mg/ml were measured at 10°C. 20 successive 0.050 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 low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Storage temperature = UNKNOWN

Tetracycline repressor (class D) (TetR(D))
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Dimer
Mon. MW   23.3 kDa
 
UniProt   P0ACT4 (2-208)
Sequence   FASTA
 
5a,6-anhydrotetracycline (ATc)
Mol. type   Other
Olig. state   Dimer
Mon. MW   0.4 kDa
Chemical formula
 
PDB ID   4D7M