HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Yan, X. Articles by Zhu, G. Search for Related Content PubMed PubMed Citation Articles by Yan, X. Articles by Zhu, G. Social Bookmarking                      What's this?

J Biol Chem, Vol. 275, Issue 10, 6712-6716, March 10, 2000

NMR Studies of Bacillus subtilis tRNA^Trp Hyperexpressed in Escherichia coli
ASSIGNMENT OF IMINO PROTON SIGNALS AND DETERMINATION OF THERMAL STABILITY^*

Xianzhong Yan, Hong Xue, Hongzhi Liu, Jun Hang, J. Tze-Fei Wong, and Guang Zhu

From the Department of Biochemistry, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China

¹⁵N-Labeled Bacillus subtilis tRNA^Trp wild type and a series of mutants were hyperexpressed in Escherichia coli and purified for NMR studies with the use of two-dimensional nuclear Overhauser effect spectroscopy (NOESY) and heteronuclear single quantum correlation (HSQC) and three-dimensional NOESY-HSQC techniques. These made possible chemical shift assignments of imino protons and determination of the thermal stability of the tRNA^Trp molecules. Almost all of the imino protons in the helical regions and the tertiary base pairs were assigned, except three imino protons of the AU base pairs whose peaks were not clearly observed. Several base triplets found in the crystal structure of tRNA were observed in the present study as well. These studies also revealed two components of tRNA^Trp, which could not be separated by high pressure liquid chromatography, corresponding to s⁴U and U at position 8 of the tRNA^Trp, as indicated by two different sets of peaks for the TC and D arms. The modification at position 8 altered the local conformation of the core region of the tRNA. Thermal unfolding experiments showed that the unfolding process is cooperative in the presence of a high concentration of magnesium ions and that the component corresponding to the s⁴U8 is more stable than the U8 component, thus providing evidence that the thiolation of U8 stabilizes the tertiary structure of tRNA.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				Q. Gong, Q. Guo, K.-L. Tong, G. Zhu, J. T.-F. Wong, and H. Xue NMR Analysis of Bovine tRNATrp. CONFORMATION DEPENDENCE OF Mg2+ BINDING J. Biol. Chem., May 31, 2002; 277(23): 20694 - 20701. [Abstract] [Full Text] [PDF]

				Q. Guo, Q. Gong, K.-L. Tong, B. Vestergaard, A. Costa, J. Desgres, M. Wong, H. Grosjean, G. Zhu, J. T.-F. Wong, et al. Recognition by Tryptophanyl-tRNA Synthetases of Discriminator Base on tRNATrp from Three Biological Domains J. Biol. Chem., April 12, 2002; 277(16): 14343 - 14349. [Abstract] [Full Text] [PDF]