Overview
- Editors:
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Joseph D. Puglisi
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SMRL & Dept. of Structural Biology D105A Fairchild Science Building, Stanford University, Stanford, USA
- Presents state of the art information on NMR spectroscopy, and its place in the broader field of biophysics
- No other monograph presents such a wide range of topics, including NMR spectroscopy, protein folding, X-ray crystallography, spectroscopy and applications
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Table of contents (8 papers)
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- Eric R. Henry, William A. Eaton
Pages 1-20
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- Timothy V. Pyrkov, Anton O. Chugunov, Nikolay A. Krylov, Dimitry E. Nolde, Roman G. Efremov
Pages 21-41
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- Saulius KlimaŠauskas, Zita LiutkeviČiŪtĖ, Dalia DaujotytĖ
Pages 51-64
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- Corey W. Liu, Viktor Y. Alekseyev, Jeffrey R. Allwardt, Alexander J. Bankovich, Barbara J. Cade-Menun, Ronald W. Davis et al.
Pages 65-81
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- Colin EcheverrÍa Aitken, R. Andrew Marshall, Joseph D. Pugi
Pages 83-99
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- Ian M. Robertson, Leo Spyracopoulos, Brian D. Sykes
Pages 101-119
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Back Matter
Pages 157-179
About this book
Single-molecule techniques eliminate ensemble averaging, thus revealing transient or rare species in heterogeneous systems [1–3]. These approaches have been employed to probe myriad biological phenomena, including protein and RNA folding [4–6], enzyme kinetics [7, 8], and even protein biosynthesis [1, 9, 10]. In particular, immobilization-based fluorescence te- niques such as total internal reflection fluorescence microscopy (TIRF-M) have recently allowed for the observation of multiple events on the millis- onds to seconds timescale [11–13]. Single-molecule fluorescence methods are challenged by the instability of single fluorophores. The organic fluorophores commonly employed in single-molecule studies of biological systems display fast photobleaching, intensity fluctuations on the millisecond timescale (blinking), or both. These phenomena limit observation time and complicate the interpretation of fl- rescence fluctuations [14, 15]. Molecular oxygen (O) modulates dye stability. Triplet O efficiently 2 2 quenches dye triplet states responsible for blinking. This results in the for- tion of singlet oxygen [16–18]. Singlet O reacts efficiently with organic dyes, 2 amino acids, and nucleobases [19, 20]. Oxidized dyes are no longer fluor- cent; oxidative damage impairs the folding and function of biomolecules. In the presence of saturating dissolved O , blinking of fluorescent dyes is sup- 2 pressed, but oxidative damage to dyes and biomolecules is rapid. Enzymatic O -scavenging systems are commonly employed to ameliorate dye instability. 2 Small molecules are often employed to suppress blinking at low O levels.
Editors and Affiliations
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SMRL & Dept. of Structural Biology D105A Fairchild Science Building, Stanford University, Stanford, USA
Joseph D. Puglisi