NMR analysis of bovine tRNATrp: conformation dependence of Mg2+ binding

The Journal of Biological Chemistry
Qingguo GongHong Xue

Abstract

NMR was used to study the solution structure of bovine tRNA(Trp) hyperexpressed in Escherichia coli. With the use of (15)N labeling and site-directed mutagenesis to assign overlapping resonances through the base pair replacement of U(71)A(2) by G(2)C(71), U(27)A(43) by G(27)C(43), and G(12)C(23) by U(12)A(23), the resonances of all 26 observable imino protons in the helical regions and in the tertiary interactions were assigned unambiguously by means of two-dimensional nuclear Overhauser effect spectroscopy and heteronuclear single quantum coherence methods. When the discriminator base A(73) and the G(12)C(23) base pair on the D stem, two identity elements on bovine tRNA(Trp) that are important for effective recognition by tryptophanyl-tRNA synthetase, were mutated to the ineffective forms of G(73) and U(12)A(23), respectively, NMR analysis revealed an important conformational change in the U(12)A(23) mutant but not in the G(73) mutant molecule. Thus A(73) appears to be directly recognized by tryptophanyl-tRNA synthetase, and G(12)C(23) represents an important structural determinant. Mg(2+) effects on the assigned resonances of imino protons allowed the identification of strong, medium, and weak Mg(2+) binding sites in tRNA(Trp...Continue Reading

References

Apr 15, 1977·Journal of Molecular Biology·A JackA Klug
Jan 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·G J QuigleyA Rich
Nov 19, 1976·Science·G J Quigley, A Rich
Feb 17, 1986·European Journal of Biochemistry·E I Hyde
Jul 11, 1989·Biochemistry·K B HallA G Redfield
Aug 16, 1974·Nature·J D RobertusA Klug
Jan 1, 1980·Annual Review of Biophysics and Bioengineering·P R Schimmel, A G Redfield
Dec 25, 1980·Nature·D MorasR Giegé
Jun 22, 1999·Journal of Molecular Biology·R L Gonzalez, I Tinoco
Jul 2, 1999·Journal of Molecular Biology·G Colmenarejo, I Tinoco
Jun 3, 2000·Journal of Molecular Biology·V K Misra, D E Draper

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.