Haloferax volcanii SILAC study of Oxidative stress

ABSTRACT: SILAC was used to monitor proteome changes to Haloferax volcanii after treatment with oxidizing agent NaOCl. SILAC amino acids used were Lysine(+8) and Arginine(+6). For treatment and control groups 4 replicates were used. In replicates 1 and 3, the control group had supplementation of both light lysine and argninie to the medium and the treatment group was supplemented with both heavy lysine(+8) and arginine(+6). In replicates 2 and 4, the labeling was switched, with the control containing heavy lysine(+8) and arginine(+6) and the treatement was supplemented with light lysine and arginine. After treatment, cells were mixed at a 1:1 ratio (control:treatment) and proteins were extracted and digested with trypsin. For the incorporation test, cells were grown in meduim containing heavy lysine(+8) and argnine(+6) and allowed to grow for 24 hours, then subcultured twice, sequentially after 24 h of growth. To test for incorporation of the heavy amino acids, cells were harvested, proteins extracted, and digested with trypsin.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Haloferax Volcanii Ds2

SUBMITTER: Lana McMillan  

LAB HEAD: Julie A. Maupin-Furlow

PROVIDER: PXD006877 | Pride | 2018-01-09

REPOSITORIES: Pride