title: Examining the molecular mechanism of bcl-2 family proteins at membranes by fluorescence spectroscopy.
abstract: The Bcl-2 family proteins control apoptosis by regulation of outer mitochondrial membrane permeabilization. Studying the Bcl-2 family is particularly difficult because the functional interactions that regulate apoptosis occur at or within intracellular membranes. Compared to other biophysical methods, fluorescence spectroscopy is well suited to study membrane-bound proteins as experiments can be performed with intact membranes and at protein concentrations similar to those found in cells. For these reasons, fluorescence spectroscopy has been particularly useful in studying the regulation of membrane permeabilization by Bcl-2 family proteins. Here, we discuss four fluorescence-based assays used to study protein dynamics at membranes, with a focus on how these techniques can be used to study the Bcl-2 family proteins.
authors: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Canada; Department of Biological Sciences, Sunnybrook Research Institute, Toronto, Canada.
authors: Justin
authors: J
authors: Kale
authors: Department of Biological Sciences, Sunnybrook Research Institute, Toronto, Canada; Department of Chemical Biology, McMaster University, Hamilton, Canada.
authors: Xiaoke
authors: X
authors: Chi
authors: Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Canada; Department of Medicine, McMaster University, Hamilton, Canada.
authors: Brian
authors: B
authors: Leber
authors: Department of Biological Sciences, Sunnybrook Research Institute, Toronto, Canada. Electronic address: david.andrews@sri.utoronto.ca.
authors: David
authors: D
authors: Andrews
mesh_terms: D000818:Animals; D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein / Q000032:analysis / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002463:Cell Membrane Permeability; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D051028:bcl-2-Associated X Protein / Q000032:analysis / Q000378:metabolism
chemical_list: D051023:BH3 Interacting Domain Death Agonist Protein; D008081:Liposomes; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein
keywords: Apoptosis; Bax; Bcl-2; Bid; Environment sensitive; FRET; Fluorescence; Membrane permeabilization; Quenching