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Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker
Published on: March 31, 2018
Temporal profiling of the adipocyte proteome during differentiation using a five-plex SILAC based strategy.
Henrik Molina1,2, Yi Yang1,2, Travis Ruch2
1McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, MD 21205, USA.
Journal of Proteome Research
|October 25, 2008
Summary
This study used a five-plex Stable Isotope Labeling with Amino acids in Cell culture (SILAC) experiment to track protein changes during adipocyte differentiation. The approach identified novel proteins involved in this crucial fat cell development process.
Area of Science:
- Proteomics
- Cell Biology
- Endocrinology
Background:
- Adipose tissue plays key roles in human endocrine and secretory functions.
- Understanding adipocyte differentiation is crucial for metabolic health.
- Transcriptomic methods have been used, but quantitative proteomics offers temporal insights.
Purpose of the Study:
- To develop and apply a five-plex SILAC experiment for temporal profiling of adipocyte differentiation.
- To investigate changes in the nuclear proteome and secretome during adipogenesis.
- To identify novel proteins differentially expressed during adipocyte differentiation.
Main Methods:
- Utilized a five-plex Stable Isotope Labeling with Amino acids in Cell culture (SILAC) approach.
- Employed four heavy stable isotopic forms of arginine for protein labeling.
- Analyzed nuclear and secretome samples using tandem mass spectrometry (quadrupole time-of-flight).
Main Results:
- Identified a total of 882 proteins from the nuclear proteome and secretome.
- Quantified 427 proteins based on arginine-containing peptides.
- Discovered novel differentially expressed proteins, including THO complex 4 and SNF2 alpha, alongside known markers like adiponectin.
Conclusions:
- The five-plex SILAC approach effectively captures dynamic proteomic changes during adipocyte differentiation.
- This method provides a powerful tool for identifying differentially expressed proteins in a temporal manner.
- The study expands the understanding of molecular regulation during adipogenesis.
