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Host cholesterol modulates the generation and enrichment of persister population during Mycobacterium tuberculosis infection.

ABSTRACT: BCG strain overexpressing the toxin (vapC12)–antitoxin (vapB12)-His(6X) locus was used for sample preparation. Log phase culture of the overexpressed strain was washed with PBS and inoculated in minimal media with 0.1% glycerol and minimal media with 0.01% cholesterol. The cultures were allowed to grow for 48 hrs and cell lysate was prepared. Immunoprecipitation was performed using an anti-His antibody (BTL1010, Biospecs). In-solution digestion was carried out for 10 µg of proteins from each condition. The samples were subjected to reduction and alkylation using 5 mM dithiothreitol (DTT) (60°C for 45 minutes) and alkylation using 10 mM Iodoacetamide (IAA). Trypsin (Gold mass-spectrometry trypsin; Promega, Madison, WI) digestion was carried out at 37°C for 10-12 hrs. The peptides were vacuum-dried and stored at -80°C until LC-MS/MS analysis.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Mycobacterium Tuberculosis (ncbitaxon:1773)

SUBMITTER: Dr. Amit Kumar Pandey

PROVIDER: MSV000086278 | MassIVE | Wed Oct 14 01:56:00 BST 2020

REPOSITORIES: MassIVE

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