Cryogenic optical localization provides 3D protein structure data with Angstrom resolution

Nat Methods. 2017 Feb;14(2):141-144. doi: 10.1038/nmeth.4141. Epub 2017 Jan 9.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Biotin / chemistry
  • Biotin / metabolism
  • Crystallography, X-Ray
  • Fluorescent Dyes / analysis*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Freezing
  • Geobacillus / chemistry
  • Histidine Kinase / chemistry*
  • Histidine Kinase / metabolism
  • Image Processing, Computer-Assisted / methods
  • Magnetic Resonance Spectroscopy
  • Microscopy, Fluorescence / methods*
  • Optics and Photonics / instrumentation
  • Optics and Photonics / methods*
  • Protein Conformation
  • Protein Domains
  • Single Molecule Imaging / methods*
  • Stochastic Processes
  • Streptavidin / metabolism

Substances

  • Bacterial Proteins
  • Fluorescent Dyes
  • Biotin
  • Streptavidin
  • Histidine Kinase