Mass spectrometry dataset for: "Discovery of Nostatin A, an azole containing sactipeptide with prominent cytostatic activity and pro-apoptotic activity"

MSn mass spectrometry dataset used in: Discovery of nostatin A, an azole-containing sactipeptide with prominent cytostatic and pro-apoptotic activity. Kateřina Delawská*, Jan Hájek*, Kateřina Voráčová*, Marek Kuzma, Jan Mareš, Kateřina Vicková, Alan Kádek, Dominika Tučková, Filip Gallob, Petra Divoká, Martin Moos, Stanislav Opekar, Lukas Koch, Kumar Saurav, David Sedlák, Petr Novák, Petra Urajová, Jason Dean, Radek Gažák, Timo J.H. Niedermeyer, Zdeněk Kameník, Petr Šimek, Andreas Villunger and Pavel Hrouzek. Org. Biomol. Chem. (2025). doi:10.1039/D4OB01395F

Description:

MSn mass spectrometry elucidation of the molecular structure of nostatin A, a bioactive peptide isolated from Nostoc sp. cyanobacteria.

Sample and data processing:

1) FTICR data:Experiments were performed using a 15T SolariX XR Fourier-transform ion cyclotron resonance mass spectrometer (ESI-FTICR MS; Bruker Daltonics, Billerica, MA, USA) equipped with infrared multiple photon dissociation (IRMPD). All data were acquired using 2 µl/min direct infusion of NosA dissolved at 10 µM in 60% acetonitrile acidified with 0.1% formic acid. Ion fragmentation was performed using IRMPD inside the ICR cell. For this a Diamond C-30A CO2 laser (Coherent, Santa Clara, CA, USA) resonating at 10.6 µm was custom-coupled to the SolariX FTICR MS and laser pulses were precisely timed in synchronization with the ICR pulse sequence. Further MS3 fragmentation experiments were performed using in source collisional fragmentation (isCID) followed by quadrupole isolation and subsequent collision induced fragmentation of particular fragment ions of interest. Detailed parameters (ESI and MS settings) are stored inside the metadata of the individual data files as well as described in the resulting publication. Spectral peaks were also exported in plain m/z vs intensity XY text files from Bruker Data Analysis 5.1.

2) qTOF data:HPLC-HRMS experiment was performed using a Dionex UltiMate 3000 HPLC system (Thermo Scientific, Sunnyvale, CA, USA) coupled with a diode array detector (DAD) connected to the Bruker Impact HD mass spectrometer equipped with an electrospray ionization (ESI) source (ESI-HRMS; Bruker, Billerica, MA, USA). The separations were performed on a C18 column (Phenomenex Kinetex C18, 150 × 4.6 mm, 2.6 μm) eluted with water (A)/acetonitrile (B) gradient (0 min 15%, 1 min 15%, 20 min 100%, 25 min 100%, 30 min 15%, 33min 15% of B) at a constant flow rate of 0.6 mL/min. Both solvents were acidified with 0.1% formic acid. Fragmentation energy was set to 87eV and 35eV for 1+ and 2+ Nostatin A, respectively. Detailed parameters (ESI and MS settings) are stored in the metadata of the individual data file as well as described in the resulting publication.

*other variants of Nostatin bearing different acyl chains on proline residue were detected in crude extract only (C33_nosA_LCMS_2217.d) and not purified.

Raw Bruker DataAnalysis .d files, which otherwise behave as folders, were compressed with the TAR algorithm implemented in the 64-bit Total Commander 11.02.