Quantifying the Assembly of Multicomponent Molecular Machines by Single-Molecule Total Internal Reflection Fluorescence Microscopy

Content Provider	Scilit
Author	Boehm, E. M. Subramanyam, S. Ghoneim, M. Washington, M. Todd Spies, M.
Copyright Year	2016
Description	Large, dynamic macromolecular complexes play essential roles in many cellular processes. Knowing how the components of these complexes associate with one another and undergo structural rearrangements is critical to understanding how they function. Single-molecule total internal reflection fluorescence (TIRF) microscopy is a powerful approach for addressing these fundamental issues. In this article, we first discuss single-molecule TIRF microscopes and strategies to immobilize and fluorescently label macromolecules. We then review the use of single-molecule TIRF microscopy to study the formation of binary macromolecular complexes using one-color imaging and inhibitors. We conclude with a discussion of the use of TIRF microscopy to examine the formation of higher-order (i.e., ternary) complexes using multicolor setups. The focus throughout this article is on experimental design, controls, data acquisition, and data analysis. We hope that single-molecule TIRF microscopy, which has largely been the province of specialists, will soon become as common in the tool box of biophysicists and biochemists as structural approaches have become today. Book Name: Protein \3- DNA Interactions
Related Links	http://europepmc.org/articles/pmc5403009?pdf=render https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5403009/pdf
Ending Page	145
Page Count	41
Starting Page	105
DOI	10.1016/bs.mie.2016.08.019
Volume Number	581
Language	English
Publisher	Elsevier BV
Publisher Date	2016-10-10
Access Restriction	Open
Subject Keyword	Book Name: Protein \3- DNA Interactions Total Internal Reflection Fluorescence Microscopy Single-molecule Fluorescence Colocalization Single-molecule Imaging Protein–protein and Protein–nucleic Acid Interaction Competitive and Noncompetitive Inhibition Kinetic Event Resolving Algorithm
Content Type	Text
Resource Type	Article