Proteomics analysis of NAD+ depletion combined with reovirus infection in KMS12 multiple myeloma cells

Published: 22 November 2021| Version 1 | DOI: 10.17632/7cty2zbz4c.1
Contributor:
Barry Kennedy

Description

Proteomics dataset of multiple myeloma cancer cell lines infected with reovirus and/or in the context of NAD+ synthesis inhibition. Experimentally, the human multiple myeloma cell line (KMS12PE) were infected or not (not-treated [NT]) with reovirus (Reo, multitude of infectivity of 10) and/or treated with NAMPT inhibitor FK866 (FK, 20 nM) and/or NAMPT product, NMN (100 uM) for 24 hours. Trypsin-digested peptides were labeled using TMT 11-plex reagents as described previously (doi.org/10.1074/mcp.M114.045849). TMT11-labeled samples were fractionated using high-pH reverse-phase chromatography performed with an Onyx monolithic 100 × 4.6-mm C18 column (Phenomenex). Fractions were desalted using homemade Stage Tips, lyophilized, and analyzed with an Orbitrap Velos Mass Spectrometer (Thermo Fisher Scientific) using an MS3 method as described previously (doi.org/10.1074/mcp.M114.045849). Protein identification was performed using a database search against a mouse proteome database (downloaded from UniProtKB in September 2014) concatenated to a mammalian orthoreovirus 3 (Dearing strain) database (downloaded from UniProtKB in September 2014). All FDR filtering and protein quantitation was performed as described previously (doi.org/10.1074/mcp.M114.045849). A protein was required to have a minimum total signal-to-noise ratio of 100 in all TMT reporter channels.

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Dalhousie University

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Proteomics

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