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. Author manuscript; available in PMC: 2021 Jan 6.
Published in final edited form as: J Proteomics. 2019 Oct 15;210:103531. doi: 10.1016/j.jprot.2019.103531

Figure 1: Workflow overview.

Figure 1:

A) overnight cultures (started from agar plates) were grown to high density in the appropriate growth media (SC for S. cerevisiae and EMM for S. pombe) with 2% pyruvate or 2% glucose. These cultures were diluted back to OD 0.1. Cells were harvested at OD 0.8 and lysed via bead beating. Fifty micrograms of cleared protein lysates were reduced and alkylated. B) The lysate was then subjected to SP3 bead-based processing to aggregate, desalt, and digest protein. C) The samples were labeled with TMT, pooled, and fractionated offline via basic pH reversed-phase (BPRP)-HPLC and desalted. Concatenated fractions (24) were analyzed on an Orbitrap Fusion Lumos mass spectrometer.