Abstract
Fractionation of cytoplasmic and nuclear proteins is a well-recognized biochemical technique to detect the intracellular distribution and expression level of proteins of interest. In the last decade, accumulating evidence shows that various types of cell surface receptors, such as receptor tyrosine kinases (RTKs), peptide hormone receptors, and cytokine receptors, are detected in the nuclei. Therefore, subcellular fractionation, including nonnuclear/nuclear extraction and the subsequent subnuclear fractionation without detectable cross-contamination during the process, is critical for studying membrane receptors that transit from the cell surface to the nucleus. Here, we utilize the epidermal growth factor receptor (EGFR) tyrosine kinase as an example of a comprehensive biochemical protocol for isolating membrane receptors in the nuclei of cancer cells.
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Acknowledgements
This study was funded in part by the following grants: National Institutes of Health (CA109311, CA099031, and CCSG CA16672); National Breast Cancer Foundation; the University of Texas MD Anderson-China Medical University and Hospital Sister Institution Fund; Cancer Research Center of Excellence (MOHW103-TD-B-111-03, Taiwan); Program for Stem Cell and Regenerative Medicine Frontier Research (NSC102-2321-B-039-001, Taiwan); International Research-Intensive Centers of Excellence in Taiwan (NSC103-2911-I-002-303, Taiwan); and the Center for Biological Pathways.
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Wang, YN., Huo, L., Hsu, J.L., Hung, MC. (2015). Biochemical Fractionation of Membrane Receptors in the Nucleus. In: Allen, B., Hébert, T. (eds) Nuclear G-Protein Coupled Receptors. Methods in Molecular Biology, vol 1234. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1755-6_9
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DOI: https://doi.org/10.1007/978-1-4939-1755-6_9
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-1755-6
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