Fluorescence applications in molecular neurobiology.
Macromolecules drive the complex behavior of neurons. For example, channels and transporters control the movements of ions across membranes, SNAREs direct the fusion of vesicles at the synapse, and motors move cargo throughout the cell. Understanding the structure, assembly, and conformational movements of these and other neuronal proteins is essential to understanding the brain. Developments in fluorescence have allowed the architecture and dynamics of proteins to be studied in real time and in a cellular context with great accuracy. In this review, we cover classic and recent methods for studying protein structure, assembly, and dynamics with fluorescence. These methods include fluorescence and luminescence resonance energy transfer, single-molecule bleaching analysis, intensity measurements, colocalization microscopy, electron transfer, and bimolecular complementation analysis. We present the principles of these methods, highlight recent work that uses the methods, and discuss a framework for interpreting results as they apply to molecular neurobiology.
Taraska
Justin W
JW
Laboratory of Molecular Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA. taraskajw@mail.nih.gov
Zagotta
William N
WN
D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D005456:Fluorescent Dyes; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D009446:Neurobiology / Q000379:methods*; D009474:Neurons / Q000502:physiology; D011487:Protein Conformation
D005456:Fluorescent Dyes