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Whole cell proteome regulation by microRNAs captured in a pulsed SILAC mass spectrometry approach

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Item Type:Article
Title:Whole cell proteome regulation by microRNAs captured in a pulsed SILAC mass spectrometry approach
Creators Name:Ebner, O.A. and Selbach, M.
Abstract:Since gene expression is controlled on many different levels in a cell, capturing a comprehensive snapshot of all regulatory processes is a difficult task. One possibility to monitor effective changes within a cell is to directly quantify changes in protein synthesis, which reflects the accumulative impact of regulatory mechanisms on gene expression. Pulsed stable isotope labeling by amino acids in cell culture (pSILAC) has been shown to be a viable method to investigate de novo protein synthesis on a proteome-wide scale (Schwanhausser et al., Proteomics 9:205-209, 2009; Selbach et al., Nature 455:58-63, 2008). One application of pSILAC is to study the regulation of protein expression by microRNAs. Here, we describe how pSILAC in conjunction with shotgun mass spectrometry can assess differences in the protein profile between cells transfected with a microRNA and non-transfected cells.
Keywords:Mass Spectrometry, LC-MS/MS, SILAC, microRNA, Seed, Transfection, In Gel Digestion
Source:Methods in Molecular Biology
Title of Book:Argonaute proteins
Publisher:Springer / Humana Press
Page Range:315-331
Additional Information:The original publication is available at www.springerlink.com
Official Publication:https://doi.org/10.1007/978-1-61779-046-1_20
PubMed:View item in PubMed

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