15N labeled e. coli tRNAMet, tRNAGlu, tRNATyr, and tRNAPhe. Double resonance and two dimensional NMR of H-N1 units in pseudouridine

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Publication Type Journal Article
School or College College of Science
Department Chemistry
Creator Poulter, Charles Dale; Davis, Darrell R.
Other Author Griffey, R. H.; Yamaizumi, Z.; Bax, A.; Nishimura, S.; Hawkins, B. L.
Title 15N labeled e. coli tRNAMet, tRNAGlu, tRNATyr, and tRNAPhe. Double resonance and two dimensional NMR of H-N1 units in pseudouridine
Date 1985-08-15
Description The N1 imino units in Escherichia coli tRNAfMet, tRNAGlu, tRNAPhe, and tRNATyr were studied by 1H-15N NMR using three different techniques to suppress signals of protons not attached to 15N. Two of the procedures, Fourier internuclear difference spectroscopy and two-dimensional forbidden echo spectroscopy permitted 1H and 15N chemical shifts to be measured simultaneously at 1H sensitivity. The tRNAs were labeled by fermentation of the uracil auxotroph S phi 187 on a minimal medium containing [1-15N]uracil. 1H and 15N resonances were detected for all of the N1 psi imino units except psi 13 at the end of the dihydrouridine stem in tRNAGlu. Chemical shifts for imino units in the tRNAs were compared with "intrinsic" values in model systems. The comparisons show that the A X psi pairs at the base of the anticodon stem in E. coli tRNAPhe and tRNATyr have psi in an anti conformation. The N1 protons of psi in other locations, including psi 32 in the anticodon loop of tRNAPhe, form internal hydrogen bonds to bridging water molecules or 2'-hydroxyl groups in nearby ribose units. These interactions permit psi to stabilize the tertiary structure of a tRNA beyond what is provided by the U it replaces.
Type Text
Publisher American Society for Biochemistry and Molecular Biology (ASBMB)
Journal Title Journal of Biological Chemistry
Volume 260
Issue 17
First Page 9734
Last Page 9741
Subject Escherichia coli; Magnetic resonance spectroscopy; Nucleic acid conformation
Subject MESH Pseudouridine; RNA, Transfer, Amino Acyl; Uridine
Language eng
Bibliographic Citation Poulter, C. D., Griffey, R. H., Davis, D., Yamaizumi, Z., Bax, A., Nishimura, S., & Hawkins, B. L. (1985). 15N labeled e. coli tRNAMet, tRNAGlu, tRNATyr, and tRNAPhe. Double resonance and two dimensional NMR of H-N1 units in pseudouridine. Journal of Biological Chemistry, 260(17), 9734-41.
Rights Management (c)American Society for Biochemistry and Molecular Biology (ASBMB)
Format Medium application/pdf
Format Extent 786,636 bytes
Identifier ir-main,1874
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Reference URL https://collections.lib.utah.edu/ark:/87278/s6bv817t