INVESTIGATING NOVEL DNA FLUORSCENCE LABELING PROBES

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DESCRIPTION (adapted from the applicant's description): The thrust of the project is to develop and employ ground-breaking methods in the use of infrared spectroscopy for the study of biological macromolecules. Developments will include infrared analogues of NMR spectroscopy for the determination of the dynamics of structure change; heterodyned 2D and 3D IR methods for study of amide I and N-H stretch modes of proteins; pump/probe methods with IR pulses, and methods to determine energy transport between remote C=O and N-H or O-H groups in peptides and proteins; methods to measure and interpret IR photon echoes and vibrational frequency correlation functions essential for understanding the response functions needed for 2D IR; instrumentation to advance T-jumps associated with IR, and of mixing kinetic methods by multiplexing infrared detection; T-jumps and mixing methods using IR probes of the intermediates; immobilization techniques to examine equilibrium fluctuations and kinetics of proteins, and other assemblies such as cells, using single-molecule fluorescence methods; methods to study ultrafast light-induced processes in biological systems probed by optical and infrared methods. There are numerous collaborative projects designed to push the technologies being developed. These involve studies on reaction centers, rhodopsin and bacteriorhodopsin, the ribosome and t-RNA, LH2 single assemblies and terahertz band-to-band transitions, pentapeptide 2D IR, calmodulin, NO-myoglobin, 310 helicies, single coiled-coils, folding and unfolding of proteins, ?-helix 2D IR, interleukin 5, ?-interferon, single ion channels, cytochrome-c trehalose, prophyrin arrays, nitrile as a probe of conformations, triplet state singles, DNA base sequence probes, and rapid-mixing folding studies. There are service projects proposed in ultrafast optical and infrared, two-dimension and single -molecule spectroscopy, in areas of DNA helicases, protein folding, serpin-serine protease interaction, laser-induced pH juimps, porphyrin electon transfer, chemosensors, helix-coil transitions, ribonucleotide reductase, light-harvesting self assembly, and NorFES single peptide study. The Resource expects to publish two newsletters each year. Besides regular seminars they will hold a synmposium on multidimensional spectroscopies and a workshop in advances in infrared laser spectroscopy . The Resource will train visiting faculty researchers, post doctorals, and graduate and undergraduate students in the applicaitons of lasers to biology.