PGF-NMR experiments were run on an 800 MHz Bruker Avance spectrometer at 25 °C. The different protein samples were prepared in D2O at a protein concentration of 50 µM in 1 mM sodium phosphate buffer (pD 6.0). The experiments were performed using the Bruker pulse sequence and the Bruker macro diffusion ordered spectroscopy (DOSY)44 (link),51 (link). Typically 16 values of gradient strength were used in the range 0 to 32 G/cm, with PFG duration of 2 ms, and diffusion time of 150 ms. The self-diffusion coefficients (Ds) were calculated using the Bruker DOSY analysis program. Each sample was run in triplicate and Ds values were averaged over the three experiments. The resulting decay curves were fitted and Ds values were calculated with the equation below:
I = I(0)exp[-D(γgδ)2(Δ-(δ/3))]
Where I(0) is 1.002, γ is 4.258 × 103 Hz/G, δ is 4.000 ms, and Δ is 150 ms.
I = I(0)exp[-D(γgδ)2(Δ-(δ/3))]
Where I(0) is 1.002, γ is 4.258 × 103 Hz/G, δ is 4.000 ms, and Δ is 150 ms.
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