Quantitative Measurements of Protein Interactions in a Crowded Cellular Environment

@article{citeulike:2959651, abstract = {Abstract: Quantitative measurements of protein interaction strengths, crucial for describing signaling networks and predicting cellular responses to environmental stimuli, are typically performed in dilute buffer solutions. However, proteinprotein interactions in cells occur within the context of a crowded system, which is characterized by a high macromolecular concentration. In this paper, we explore the utility of cell-derived vesicles as a model crowded environment for quantitative FRET measurements of proteinprotein interactions. We show that the FRET efficiency, and the donor and acceptor concentrations, can be calculated in each vesicle. We also introduce the quantitative imaging Foster resonance energy transfer method as a tool that can yield protein interaction strengths within these vesicles.}, address = {Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland 21218}, author = {Li, Edwin and Placone, Jesse and Merzlyakov, Mikhail and Hristova, Kalina }, citeulike-article-id = {2959651}, doi = {http://dx.doi.org/10.1021/ac800616u}, journal = {Anal. Chem.}, keywords = {fret, interaction, proteomics}, month = {July}, posted-at = {2008-07-03 16:07:32}, priority = {2}, title = {Quantitative Measurements of Protein Interactions in a Crowded Cellular Environment}, url = {http://dx.doi.org/10.1021/ac800616u}, year = {2008} }

TY - JOUR ID - citeulike:2959651 L3 - citeulike-article-id:2959651 TI - Quantitative Measurements of Protein Interactions in a Crowded Cellular Environment JF - Anal. Chem. AD - Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland 21218 N2 - Abstract: Quantitative measurements of protein interaction strengths, crucial for describing signaling networks and predicting cellular responses to environmental stimuli, are typically performed in dilute buffer solutions. However, proteinprotein interactions in cells occur within the context of a crowded system, which is characterized by a high macromolecular concentration. In this paper, we explore the utility of cell-derived vesicles as a model crowded environment for quantitative FRET measurements of proteinprotein interactions. We show that the FRET efficiency, and the donor and acceptor concentrations, can be calculated in each vesicle. We also introduce the quantitative imaging Foster resonance energy transfer method as a tool that can yield protein interaction strengths within these vesicles. KW - fret KW - interaction KW - proteomics AU - Li, Edwin AU - Placone, Jesse AU - Merzlyakov, Mikhail AU - Hristova, Kalina PY - 2008/07/03/ UR - http://dx.doi.org/10.1021/ac800616u ER -