posted on 2024-10-18, 22:18authored byEsther ShmidovEsther Shmidov, Alexis Villani, Joseph Bondy-Denomy, Ehud Banin
<p dir="ltr">The data in this item includes raw proteomics data from post-43°C exposure and during the recovery period to assess protein-level effect.</p><p dir="ltr"><b>Sample preparation</b>: Overall, 18 samples including 3 replicates of <i>Pseudomonas aeruginosa</i> after exposure to 37<i> </i>°C (control) or 43<i> </i>°C (heat shock), at 3 time points (T=0hr, T=18hr, and T=54hr). Samples from the T=0 groups are simply labeled “37” or “43”.The samples were subjected in-solution tryptic digestion using suspension trapping (S-trap method by Protifi).</p><p dir="ltr"><b>Liquid chromatography mass spectrometry</b>: The resulting peptides were analyzed using nanoflow liquid chromatography (nanoAcquity) coupled to high resolution, high mass accuracy mass spectrometry (Q-Exactive HF). Each sample was analyzed on the instrument separately in a random order in discovery mode.</p><p dir="ltr"><b>Data processing</b>: Raw data was processed with MaxQuant v1.6.6.0. The data was searched with the Andromeda search engine against the Pseudomonas aeruginosa PAO1 database as downloaded from Uniprot, appended with common lab protein contaminants. Search parameters included the following modifications: Fixed modification- cysteine carbamidomethylation. Variable modifications- methionine oxidation. The quantitative comparisons were calculated using Perseus v1.6.0.7. Decoy hits were filtered out and only proteins that were identified in at least two replicates of at least one experimental group were kept.</p>