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K48-Ubiquitin IP-MS of Jurkat leukemia cells treated with asparaginase

ABSTRACT: We have shown that drug-resistant leukemias rely on protein degradation through the ubiquitin proteasome pathway to survive during asparaginase therapy. Our studies support the model that this catabolic source of amino acids is upregulated in response to amino acid starvation. To investigate whether this involves bulk protein degradation or the degradation of specific protein targets, we analyzed K48-ubiquitinated proteins using immunoprecipitation with an anti-K48-linked ubiquitin antibody (Millipore #05-1307) followed by mass spectrometry proteomics from Jurkat cells after treatment with vehicle vs asparaginase. We found no enrichment of specific proteins after asparaginase treatment, consistent with the possibility that asparaginase therapy stimulates bulk protein degradation, although these results do not rule out degradation of specific proteins below the limit of detection of this assay.

INSTRUMENT(S): LTQ Orbitrap

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

SUBMITTER: Alejandro Gutierrez

PROVIDER: MSV000089226 | MassIVE | Fri Apr 08 11:57:00 BST 2022

SECONDARY ACCESSION(S): PXD033118

REPOSITORIES: MassIVE

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