Abstract
We introduce Cryogenic Optical Localization in 3D (COLD), a method to localize multiple fluorescent sites within a single small protein with Angstrom resolution. We demonstrate COLD by determining the conformational state of the cytosolic Per-ARNT-Sim domain from the histidine kinase CitA of Geobacillus thermodenitrificans and resolving the four biotin sites of streptavidin. COLD provides quantitative 3D information about small- to medium-sized biomolecules on the Angstrom scale and complements other techniques in structural biology.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacterial Proteins / chemistry
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Bacterial Proteins / metabolism
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Binding Sites
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Biotin / chemistry
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Biotin / metabolism
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Crystallography, X-Ray
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Fluorescent Dyes / analysis*
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Fluorescent Dyes / chemistry
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Fluorescent Dyes / metabolism
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Freezing
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Geobacillus / chemistry
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Histidine Kinase / chemistry*
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Histidine Kinase / metabolism
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Image Processing, Computer-Assisted / methods
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Magnetic Resonance Spectroscopy
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Microscopy, Fluorescence / methods*
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Optics and Photonics / instrumentation
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Optics and Photonics / methods*
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Protein Conformation
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Protein Domains
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Single Molecule Imaging / methods*
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Stochastic Processes
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Streptavidin / metabolism
Substances
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Bacterial Proteins
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Fluorescent Dyes
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Biotin
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Streptavidin
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Histidine Kinase