Ft nmr avance 3

Reagents were purchased from Aldrich unless specified otherwise. All solvents were purified and degassed prior to use. NMR spectra were recorded on a Bruker FT-NMR Avance III 500 MHz instrument at 500.32 (¹H), 125.81 (¹³C) MHz. Two-dimensional NMR measurements were carried out using standard pulse programs. Chemical shifts were referenced relative to the solvent signal for ¹H and ¹³C spectra. ESI mass spectra were recorded on a LC/MSn ion trap mass spectrometer amaZon SL (Bruker, Bremen, Germany) with MeOH as a solvent. Elemental analysis was performed at Moscow State University with the MicroCube Elementar analyser. Melting points were determined with a Stuart Scientific SMP3 apparatus and uncorrected. UV-vis spectra were recorded on Thermo Scientific Evolution 300. Source of cell lines: initially, all cell lines were purchased from ATCC (Manassas, VA, USA) and routinely propagated by the authors according to the manufacturer’s protocols.

We incubated a sample of 15^N labelled aS (160.5 μM) produced as previously described40 (link) in the presence of an equimolar amount of DOPAL (160.5 μM) in PBS (pH 7.4) at room temperature for 30 days. The reaction was monitored by recording 15N-1H HSQC spectra of the mixture at 25 °C. The final spectra reported are the overlap of the 15N-1H HSQC spectra of aS sample in the absence of DOPAL and after 30 days of incubation with DOPAL at 10 °C. The spectra were recorded at lower temperature than the incubation temperature due to resolution issues. The NMR experiments were recorded with a Bruker FT NMR Avance III 600-MHz spectrometer equipped with a 5-mm CryoProbe QCI 1 H/19F-13C/15N-D quadruple resonance, and a shielded z-gradient coil. The analysis of the spectra was performed using the CcpNMR software and the NMR assignment published in the BMRB (bmr16300.str) for the intrinsically unfolded aS.