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Heavy-methyl SILAC of ribosomal proteins

ABSTRACT: Methylation on ribosome subunits and ribosime-associating proteins were compared in a low- and high-SAM conditions using heavy-methyl SILAC. Ribosome proteins were isolated in two ways; one was ultracentrifugation and the other was anti-FLAG immunoprecipitation using HeLa cells transfected with a FLAG-RPS2 expression vector or FLAG-RPL23a expression vector. The cells were grown in medium containing either light- or heavy-labelled methionine and then cycloleucine, an inhibitor of SAM synthesis enzymes, was added to the heavy-labelled culture to reduce intracellular SAM level. Ribosome proteins were purified by either ultracentrifugation or anti-FLAG immunoprecipitation. he samples were separated by SDS-PAGE and were in-gel-digested with trypsin. Files 201211-1 to 4: results from ultracentrifugation, 201219-7 to 9: anti-FLAG IP using FLAG-RPL23a expressing HeLa cells.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Kazuhiko IGARASHI

PROVIDER: MSV000089296 | MassIVE | Thu Apr 21 19:46:00 BST 2022

REPOSITORIES: MassIVE

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