LC-MS/MS data for protein band

Description

The prominent band induced by growth on L-cysteate and pyruvate was manually excised. After in-gel digestion with trypsin and extraction, the mixture of peptides was loaded onto Orbitrap Fusion LUMOS MS. The MS/MS spectra from each LC-MS/MS run were searched against the B. wadsworthia protein database GCF_000185705.2 (Bilo_- wads_3_1_6_V2) from UniProt (release date of Mar, 2022; 68,406 entries) using an in-house Proteome Discoverer (Version PD 2.2, Thermo-Fisher Scientific). Protein identifications were performed based on Sequest HT.

Steps to reproduce

B. wadsworthia cells were boiled in Laemmli loading buffer for cell lysis and proteins were separated on a 10% SDS/PAGE gel which was stained with Coomassie Blue R-250 staining solution, followed by a destaining process. The most prominent band induced by growth on L-cysteate and pyruvate was manually excised. After in-gel digestion with trypsin and extraction, the mixture of peptides was loaded onto Orbitrap Fusion LUMOS MS equipped with a 150 μm × 2 cm self-packed C18 trap column (particle size 3 μm, Dr. MASCH GmbH, Germany) and a 150 μm × 30 cm self-packed C18 analytical column (particle size 1.9 μm, Dr. MASCH GmbH). The HPLC conditions were as follows: linear gradient from 8 to 12% mobile phase B over 10 min, 12 to 27% over 69 min, 27 to 45% over 28 min, 45 to 95% over 3 min, and 95% over 10 min, where mobile phase A was ddH2O and 0.1% formic acid, and mobile phase B was 80% acetonitrile and 0.1% formic acid. The MS/MS spectra from each LC-MS/MS run were searched against the B. wadsworthia protein database GCF_000185705.2 (Bilo_- wads_3_1_6_V2) from UniProt (release date of March 2022; 68,406 entries) using an in-house Proteome Discoverer (Version PD 2.2, ThermoFisher Scientific). Protein identification was performed based on Sequest HT.

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