5-fluoro-tryptophan as a probe for fluorescence and fluorine-19 NMR structure-function studies: Analysis of 5-fluoro-tryptophan substituted soluble tissue factor.
Item
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Title
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5-fluoro-tryptophan as a probe for fluorescence and fluorine-19 NMR structure-function studies: Analysis of 5-fluoro-tryptophan substituted soluble tissue factor.
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Identifier
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AAI9820597
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identifier
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9820597
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Creator
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Zemsky, Jennifer.
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Contributor
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Adviser: J. B. A. Ross
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Date
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1998
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Language
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English
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Publisher
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City University of New York.
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Subject
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Chemistry, Biochemistry | Biophysics, General
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Abstract
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Both protein fluorescence and {dollar}\sp{19}{dollar}F NMR spectroscopies are highly sensitive tools for probing the local environments of proteins. However, their usefulness becomes limited in complex multi-component systems because the components become indistinguishable. One approach to characterization of multi-protein or protein-DNA systems is to label one component such that it can be selectively observed. Proteins may be labeled either intrinsically or extrinsically. Intrinsic probes have two advantages over extrinsic probes. First, intrinsic probes are site specific. Extrinsic probes often label proteins randomly. Second, intrinsic probes often perturb protein structure less than extrinsic probes. Tryptophan analogs have proven to be useful intrinsic probes both for fluorescence and {dollar}\sp{19}{dollar}F NMR spectroscopies. Substitution on the indole ring of tryptophan can result in altered spectroscopic properties. In this study, the utility of 5-fluoro-tryptophan as an intrinsic probe has been characterized. 5-fluoro-tryptophan has two advantages as a probe. First, 5-fluoro-tryptophan may be used as both a {dollar}\sp{19}{dollar}F NMR and fluorescence probe, allowing complementary data to be gathered on the same sample by both techniques, Second, 5-fluoro-tryptophan perturbs protein structure minimally. In this study, 5-fluoro-tryptophan was substituted for tryptophan in soluble human tissue factor and its single tryptophan replacement mutants. Both the {dollar}\sp{19}{dollar}F NMR and fluorescence spectra of these proteins have been characterized. The data on the local environment has been characterized with respect to previous studies characterizing the tryptophan fluorescence of these proteins.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
