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Targeted Proteomics-Driven Computational Modeling of Macrophage S1P Chemosensing.

ABSTRACT: Osteoclasts are monocyte-derived multinuclear cells that directly attach to and resorb bone. Sphingosine-1-phosphate (S1P)(1) regulates bone resorption by functioning as both a chemoattractant and chemorepellent of osteoclast precursors through two G-protein coupled receptors that antagonize each other in an S1P-concentration-dependent manner. To quantitatively explore the behavior of this chemosensing pathway, we applied targeted proteomics, transcriptomics, and rule-based pathway modeling using the Simmune toolset. RAW264.7 cells (a mouse monocyte/macrophage cell line) were used as model osteoclast precursors, RNA-seq was used to identify expressed target proteins, and selected reaction monitoring (SRM) mass spectrometry using internal peptide standards was used to perform absolute abundance measurements of pathway proteins. The resulting transcript and protein abundance values were strongly correlated. Measured protein abundance values, used as simulation input parameters, led to in silico pathway behavior matching in vitro measurements. Moreover, once model parameters were established, even simulated responses toward stimuli that were not used for parameterization were consistent with experimental findings. These findings demonstrate the feasibility and value of combining targeted mass spectrometry with pathway modeling for advancing biological insight.

SUBMITTER: Manes NP

PROVIDER: S-EPMC4597143 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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Targeted Proteomics-Driven Computational Modeling of Macrophage S1P Chemosensing.

Manes Nathan P NP Angermann Bastian R BR Koppenol-Raab Marijke M An Eunkyung E Sjoelund Virginie H VH Sun Jing J Ishii Masaru M Germain Ronald N RN Meier-Schellersheim Martin M Nita-Lazar Aleksandra A

Molecular & cellular proteomics : MCP 20150721 10

Osteoclasts are monocyte-derived multinuclear cells that directly attach to and resorb bone. Sphingosine-1-phosphate (S1P)(1) regulates bone resorption by functioning as both a chemoattractant and chemorepellent of osteoclast precursors through two G-protein coupled receptors that antagonize each other in an S1P-concentration-dependent manner. To quantitatively explore the behavior of this chemosensing pathway, we applied targeted proteomics, transcriptomics, and rule-based pathway modeling usin ...[more]

PMID: 26199343

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Dataset Information

Targeted Proteomics-Driven Computational Modeling of Macrophage S1P Chemosensing.

Publications

Targeted Proteomics-Driven Computational Modeling of Macrophage S1P Chemosensing.

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