College of Pharmacy; College of Science; School of Medicine
Department
Biochemistry; Medicinal Chemistry; Chemistry
citation_author
Poulter, Charles Dale; Davis, Darrell R.
citation_other_author
Griffey, Richard H.; Yamaizumi, Ziro; Nishimura, Susumu; Bax, Ad; Hawkins, Bruce
citation_title
15N-labeled Escherichia coli tRNAfMet, tRNAGlu, tRNATyr, and tRNAPhe. Double resonance and two-dimensional NMR of N1-labeled pseudouridine.
citation_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
citation_publisher
American Society for Biochemistry and Molecular Biology (ASBMB)
citation_journal_title
Journal of Biological Chemistry
citation_volume
260
citation_issue
17
citation_firstpage
9734
Citation_lastpage
9741
citation_keywords
Escherichia coli; Magnetic resonance spectroscopy; Nitrogen; Nucleic acid conformation
Journal of Biological Chemistry, 1985 Aug 15, 260(17), 9734-41. Griffey RH, Davis D, Yamaizumi Z, Nishimura S, Bax A, Hawkins B, Poulter CD. 15N-labeled Escherichia coli tRNAfMet, tRNAGlu, tRNATyr, and tRNAPhe. Double resonance and two-dimensional NMR of N1-labeled pseudouridine. Retrieved April 16, 2007 from http://www.jbc.org/cgi/reprint/260/17/9734.
