Biophysics and the Challenges of Emerging Threats

2009
ISBN: 978-90-481-2366-7
Verlag: Springer Netherlands

Buch, Englisch, 179 Seiten, Format (B × H): 160 mm x 241 mm, Gewicht: 453 g

Reihe: NATO Science for Peace and Security Series B: Physics and Biophysics

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.

Zielgruppe

Research

Autoren/Hrsg.

Puglisi, Joseph

Fachgebiete

• Naturwissenschaften Biowissenschaften Angewandte Biologie Biophysik
• Naturwissenschaften Physik Thermodynamik Festkörperphysik, Kondensierte Materie
• Naturwissenschaften Biowissenschaften Biochemie (nichtmedizinisch)
• Technische Wissenschaften Verfahrenstechnik | Chemieingenieurwesen | Biotechnologie Biotechnologie Industrielle Biotechnologie
• Naturwissenschaften Physik Angewandte Physik Biophysik
• Naturwissenschaften Chemie Physikalische Chemie Quantenchemie, Theoretische Chemie

Inhaltsverzeichnis

A Simple Model for Protein Folding.- Complementarity of Hydrophobic/Hydrophilic Properties In Protein—Ligand Complexes: A New Tool to Improve Docking Results.- Structures of Cvnh Family Lectins.- Biophysical Approaches To Study Dna Base Flipping.- The Diversity of Nuclear Magnetic Resonance Spectroscopy.- Improved Dye Stability in Single-Molecule Fluorescence Experiments.- The Evaluation of Isotope Editing and Filtering for Protein—Ligand Interaction Elucidation by Nmr.- Ribosome: an Ancient Cellular Nano-Machine for Genetic Code Translation.