Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred

IMPACT FACTOR 2017: 3.022

CiteScore 2017: 2.81

SCImago Journal Rank (SJR) 2017: 1.562
Source Normalized Impact per Paper (SNIP) 2017: 0.705

See all formats and pricing
More options …
Volume 388, Issue 5


Characterization of peptidyl-tRNA hydrolase encoded by open reading frame Rv1014c of Mycobacterium tuberculosis H37Rv

Naresh C. Bal
  • 1Molecular and Structural Biology, Central Drug Research Institute, Lucknow 226 001, India
    The first two authors contributed equally to this work.
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Himanshu Agrawal / Akshaya K. Meher / Ashish Arora
Published Online: 2007-05-01 | DOI: https://doi.org/10.1515/BC.2007.057


The enzyme peptidyl-tRNA hydrolase (Pth, EC is essential for the viability of bacteria. The ORF Rv1014c of Mycobacterium tuberculosis H37Rv, designated as the mtpth gene, was cloned and over-expressed and the product was purified. Generation of polyclonal antibodies against the purified recombinant protein, termed MtPth, facilitated detection of endogenously expressed MtPth in M. tuberculosis H37Rv cell lysate. MtPth could release diacetyl-[3H]-lysine from diacetyl-[3H]-lysyl-tRNALys with Michaelis-Menten kinetic parameters of K m=0.7±0.2 μM and k cat=1.22±0.2 s-1. Transformation of a pTrc99c/mtpth vector allowed the growth of E. coli thermosensitive Pth(ts) mutant strain AA7852 at the non-permissive temperature of 42°C, demonstrating the in vivo activity of MtPth. In addition, at 39°C, over-expression of MtPth in AA7852 cells allowed the cells to remain viable in the presence of up to 200 μg/ml erythromycin. A 3D fold based on NMR and a structural model based on the E. coli Pth crystal structure were generated for MtPth. The essential nature of conserved active-site residues N12, H22 and D95 of MtPth for catalysis was demonstrated by mutagenesis and complementation in E. coli mutant strain AA7852. Thermal and urea/guanidinium chloride (GdmCl)-induced unfolding curves for MtPth indicate a simple two-state unfolding process without any intermediates.

Keywords: complementation; enzyme activity; molecular modeling; mutagenesis; peptidyl-tRNA hydrolase; thermal unfolding

About the article

Corresponding author

Received: 2006-11-02

Accepted: 2007-02-13

Published Online: 2007-05-01

Citation Information: Biological Chemistry, Volume 388, Issue 5, Pages 467–479, ISSN (Online) 14316730, ISSN (Print) 14374315, DOI: https://doi.org/10.1515/BC.2007.057.

Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

Ami Matsumoto, Yuji Uehara, Yoshihiro Shimizu, Takuya Ueda, Toshio Uchiumi, and Kosuke Ito
Proteins: Structure, Function, and Bioinformatics, 2018
Salman Shahid, Ashish Kabra, Surbhi Mundra, Ravi Kant Pal, Sarita Tripathi, Anupam Jain, and Ashish Arora
Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 2018
Naresh C. Bal, Nivedita Jena, Harapriya Chakravarty, Amit Kumar, Mei Chi, Tuniki Balaraju, Sharad V. Rawale, Jayashree S. Rawale, Ashoke Sharon, Muthu Periasamy, and Stephen C. Blacklow
Biopolymers, 2015, Volume 103, Number 1, Page 15
Laurent Giorgi, Pierre Plateau, Gavin O'Mahony, Caroline Aubard, Michel Fromant, Aurlien Thureau, Morten Grtli, Sylvain Blanquet, and Franois Bontems
Journal of Molecular Biology, 2011, Volume 412, Number 4, Page 619
S.V.S.R.K. Pulavarti, Anupam Jain, Prem Prakash Pathak, Anjum Mahmood, and Ashish Arora
Journal of Molecular Biology, 2008, Volume 378, Number 1, Page 165

Comments (0)

Please log in or register to comment.
Log in