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Single-molecule fluorescence studies from a bioinorganic perspective

Inorganica Chimica Acta
Publication Date
DOI: 10.1016/j.ica.2007.08.012
  • Single-Molecule Fluorescence Spectroscopy
  • Bioinorganic Chemistry
  • Fret
  • Fluorescence Quenching Via Energy Transfer
  • Fluorescence Quenching Via Electron Transfer
  • Oxygen Binding
  • Redox
  • Metalloprotein Folding
  • Conformational Dynamics
  • Catalysis
  • Metallochaperones
  • Metalloregulators
  • Biology
  • Chemistry


Abstract In recent years, single-molecule methods have enabled many innovative studies in the life sciences, which generated unprecedented insights into the workings of many macromolecular machineries. Single-molecule studies of bioinorganic systems have been limited, however, even though bioinorganic chemistry represents one of the frontiers in the life sciences. With the hope to stimulate more interest in applying existing and developing new single-molecule methods to address compelling bioinorganic problems, this review discusses a few single-molecule fluorescence approaches that have been or can be employed to study the functions and dynamics of metalloproteins. We focus on their principles, features and generality, possible further bioinorganic applications, and experimental challenges. The fluorescence quenching via energy transfer approach has been used to study the O 2-binding of hemocyanin, the redox states of azurin, and the folding dynamics of cytochrome c at the single-molecule level. Possible future applications of this approach to single-molecule studies of metalloenzyme catalysis and metalloprotein folding are discussed. The fluorescence quenching via electron transfer approach can probe the subtle conformational dynamics of proteins, and its possible application to probe metalloprotein structural dynamics is discussed. More examples are presented in using single-molecule fluorescence resonance energy transfer to probe metallochaperone protein interactions and metalloregulator–DNA interactions on a single-molecule basis.

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