2026-09-20T17:28:13Zhttps://dspace.mit.edu/server/oai/requestoai:dspace.mit.edu:1721.1/408742022-01-13T07:54:21Zcom_1721.1_7582com_1721.1_7581col_1721.1_131022
Robert G. Griffin.
Bajaj, Vikram Singh
Massachusetts Institute of Technology. Dept. of Chemistry.
Massachusetts Institute of Technology. Department of Chemistry
2008-03-27T18:16:37Z
2008-03-27T18:16:37Z
2007
2007
http://hdl.handle.net/1721.1/40874
181591869
Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Chemistry, 2007.
Includes bibliographical references.
Solid state NMR can probe structure and dynamics on length scales from the atomic to the supramolecular. However, low sensitivity limits its application in macromolecules. NMR sensitivity can be improved by dynamic nuclear polarization (DNP), in which electron polarization is transferred to nuclei. We present applications of magic angle spinning NMR that demonstrate its utility for the determination of structure at atomic resolution. We then present new techniques and instrumentation for DNP that permit these methods to be applied to larger systems such as membrane proteins. These applications rest on several advances in instrumentation: millimeter-wave sources and conduits of power to the sample; low-temperature MAS probes incorporating millimeter-wave transmission; cryogenics and pneumatic control systems. We describe a 380 MHz DNP spectrometer incorporating a 250 GHz gyrotron oscillator and present the theory and operation of a 460 GHz gyrotron at the second harmonic of electron cyclotron resonance. We have applied DNP to study trapped photo cycle intermediates of the archael membrane protein bacteriorhodopsin, a light-driven transmembrane ion pump.
(cont.) We have observed the K photointermediate for the first time by NMR and found unexpected conformational heterogeneity in the L intermediate. With multidimensional correlation spectroscopy, we have assigned active site resonances in conformational mixtures of photointermediates of [U-13C,'SN]-bR with high sensitivity. By using non-linear sampling of indirect dimensions, we have observed transient product of K accumulation. We present frequency-selective experiments for amino acid-selective assignments and the measurement of heteronuclear distances and torsion angles in [U-13C, N]-bR and discuss the relevance of these results to its photocycle. In addition, we describe several applications of solid state NMR, including a study of dynamic and structural phase transitions in peptides and proteins near the canonical glass transition temperature. We present resonance width experiments that can be used to measure homonuclear and heteronuclear dipolar couplings in uniformly labeled solids.
(cont.) Finally, we discuss applications to amyloid fibrils, which are protein aggregates that are implicated in diseases of protein misfolding. We report the atomic resolution structure of the disease-associated L 111M mutant of TTR105-115 in an amyloid fibril, and information about the supramolecular structure of fibrils from WT TTRos05115.
by Vikram Singh Bajaj.
Ph.D.
603, [1] p.
eng
Massachusetts Institute of Technology
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http://dspace.mit.edu/handle/1721.1/7582
Chemistry.
Dynamic nuclear polarization in biomolecular solid state NMR : methods and applications in peptides and membrane proteins
Thesis
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Publication
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<Abstract>Solid state NMR can probe structure and dynamics on length scales from the atomic to the supramolecular. However, low sensitivity limits its application in macromolecules. NMR sensitivity can be improved by dynamic nuclear polarization (DNP), in which electron polarization is transferred to nuclei. We present applications of magic angle spinning NMR that demonstrate its utility for the determination of structure at atomic resolution. We then present new techniques and instrumentation for DNP that permit these methods to be applied to larger systems such as membrane proteins. These applications rest on several advances in instrumentation: millimeter-wave sources and conduits of power to the sample; low-temperature MAS probes incorporating millimeter-wave transmission; cryogenics and pneumatic control systems. We describe a 380 MHz DNP spectrometer incorporating a 250 GHz gyrotron oscillator and present the theory and operation of a 460 GHz gyrotron at the second harmonic of electron cyclotron resonance. We have applied DNP to study trapped photo cycle intermediates of the archael membrane protein bacteriorhodopsin, a light-driven transmembrane ion pump.</Abstract>
<Abstract>(cont.) We have observed the K photointermediate for the first time by NMR and found unexpected conformational heterogeneity in the L intermediate. With multidimensional correlation spectroscopy, we have assigned active site resonances in conformational mixtures of photointermediates of [U-13C,'SN]-bR with high sensitivity. By using non-linear sampling of indirect dimensions, we have observed transient product of K accumulation. We present frequency-selective experiments for amino acid-selective assignments and the measurement of heteronuclear distances and torsion angles in [U-13C, N]-bR and discuss the relevance of these results to its photocycle. In addition, we describe several applications of solid state NMR, including a study of dynamic and structural phase transitions in peptides and proteins near the canonical glass transition temperature. We present resonance width experiments that can be used to measure homonuclear and heteronuclear dipolar couplings in uniformly labeled solids.</Abstract>
<Abstract>(cont.) Finally, we discuss applications to amyloid fibrils, which are protein aggregates that are implicated in diseases of protein misfolding. We report the atomic resolution structure of the disease-associated L 111M mutant of TTR105-115 in an amyloid fibril, and information about the supramolecular structure of fibrils from WT TTRos05115.</Abstract>
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