title: Single molecule optical measurements of orientation and rotations of biological macromolecules.
abstract: Subdomains of macromolecules often undergo large orientation changes during their catalytic cycles that are essential for their activity. Tracking these rearrangements in real time opens a powerful window into the link between protein structure and functional output. Site-specific labeling of individual molecules with polarized optical probes and measurement of their spatial orientation can give insight into the crucial conformational changes, dynamics, and fluctuations of macromolecules. Here we describe the range of single molecule optical technologies that can extract orientation information from these probes, review the relevant types of probes and labeling techniques, and highlight the advantages and disadvantages of these technologies for addressing specific inquiries.
authors.affiliation: These authors contributed equally to this work.
authors.forename: Deborah Y
authors.initials: DY
authors.lastname: Shroder
authors.forename: Lisa G
authors.initials: LG
authors.lastname: Lippert
authors.forename: Yale E
authors.initials: YE
authors.lastname: Goldman
mesh_terms: D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D061848:Optical Imaging; D052898:Optical Tweezers; D011487:Protein Conformation; D011506:Proteins; D012399:Rotation; D000072760:Single Molecule Imaging
chemical_list: D005456:Fluorescent Dyes; D046911:Macromolecular Substances; D011506:Proteins