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Pattern Analysis of Organellar Maps for Interpretation of Pro-teomic Data

ABSTRACT: The human proton-coupled folate transporter (PCFT) was stably expressed as a fu-sion protein with a BioID2-HA in the C-terminal position (PCFT-BioID2). The uncom-plxed biotin transferase was used as a control to identify proteins that are biotinylated randomly due to sheer abundance and/or an unnatural affinity to the BioID2 ligase. HeLa PCFT-BioID2 cells expressed the PCFT fusion protein which localized to the plasma mem-brane and exhibited robust transport typical of PCFT. HeLa PCFT-BioID2 and BioID2 cells were cultured in complete DMEM (biotin-free), with and without biotin (50 μM; 16 h) then biotinylated proteins were selected on streptavidin beads.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Jordan Burton

LAB HEAD: Paul M. Stemmer

PROVIDER: PXD032297 | Pride | 2022-06-09

REPOSITORIES: Pride

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Pattern Analysis of Organellar Maps for Interpretation of Proteomic Data.

Burton Jordan B JB Carruthers Nicholas J NJ Hou Zhanjun Z Matherly Larry H LH Stemmer Paul M PM

Proteomes 20220523 2

Localization of organelle proteins by isotope tagging (LOPIT) maps are a coordinate-directed representation of proteome data that can aid in biological interpretation. Analysis of organellar association for proteins as displayed using LOPIT is evaluated and interpreted for two types of proteomic data sets. First, test and control group protein abundances and fold change data obtained in a proximity labeling experiment are plotted on a LOPIT map to evaluate the likelihood of true protein interact ...[more]

PMID: 35645376

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