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Title: A functional role of Rv1738 in Mycobacterium tuberculosis persistence suggested by racemic protein crystallography

Abstract

Racemic protein crystallography was used to determine the X-ray structure of the predicted Mycobacterium tuberculosis protein Rv1738, which had been completely recalcitrant to crystallization in its natural L-form. Native chemical ligation was used to synthesize both L-protein and D-protein enantiomers of Rv1738. Crystallization of the racemic {D-protein + L-protein} mixture was immediately successful. The resulting crystals diffracted to high resolution and also enabled facile structure determination because of the quantized phases of the data from centrosymmetric crystals. The X-ray structure of Rv1738 revealed striking similarity with bacterial hibernation factors, despite minimal sequence similarity. As a result, we predict that Rv1738, which is highly up-regulated in conditions that mimic the onset of persistence, helps trigger dormancy by association with the bacterial ribosome.

Authors:
 [1];  [2];  [1];  [1];  [2];  [1];  [2];  [1]
  1. Univ. of Auckland, Auckland (New Zealand)
  2. Univ. of Chicago, Chicago, IL (United States)
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1178127
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 112; Journal Issue: 14; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; racemic protein crystallography; Mycobacterium tuberculosis; ribosome binding; dormancy; protein structure

Citation Formats

Bunker, Richard D., Mandal, Kalyaneswar, Bashiri, Ghader, Chaston, Jessica J., Pentelute, Bradley L., Lott, J. Shaun, Kent, Stephen B. H., and Baker, Edward N. A functional role of Rv1738 in Mycobacterium tuberculosis persistence suggested by racemic protein crystallography. United States: N. p., 2015. Web. doi:10.1073/pnas.1422387112.
Bunker, Richard D., Mandal, Kalyaneswar, Bashiri, Ghader, Chaston, Jessica J., Pentelute, Bradley L., Lott, J. Shaun, Kent, Stephen B. H., & Baker, Edward N. A functional role of Rv1738 in Mycobacterium tuberculosis persistence suggested by racemic protein crystallography. United States. https://doi.org/10.1073/pnas.1422387112
Bunker, Richard D., Mandal, Kalyaneswar, Bashiri, Ghader, Chaston, Jessica J., Pentelute, Bradley L., Lott, J. Shaun, Kent, Stephen B. H., and Baker, Edward N. 2015. "A functional role of Rv1738 in Mycobacterium tuberculosis persistence suggested by racemic protein crystallography". United States. https://doi.org/10.1073/pnas.1422387112. https://www.osti.gov/servlets/purl/1178127.
@article{osti_1178127,
title = {A functional role of Rv1738 in Mycobacterium tuberculosis persistence suggested by racemic protein crystallography},
author = {Bunker, Richard D. and Mandal, Kalyaneswar and Bashiri, Ghader and Chaston, Jessica J. and Pentelute, Bradley L. and Lott, J. Shaun and Kent, Stephen B. H. and Baker, Edward N.},
abstractNote = {Racemic protein crystallography was used to determine the X-ray structure of the predicted Mycobacterium tuberculosis protein Rv1738, which had been completely recalcitrant to crystallization in its natural L-form. Native chemical ligation was used to synthesize both L-protein and D-protein enantiomers of Rv1738. Crystallization of the racemic {D-protein + L-protein} mixture was immediately successful. The resulting crystals diffracted to high resolution and also enabled facile structure determination because of the quantized phases of the data from centrosymmetric crystals. The X-ray structure of Rv1738 revealed striking similarity with bacterial hibernation factors, despite minimal sequence similarity. As a result, we predict that Rv1738, which is highly up-regulated in conditions that mimic the onset of persistence, helps trigger dormancy by association with the bacterial ribosome.},
doi = {10.1073/pnas.1422387112},
url = {https://www.osti.gov/biblio/1178127}, journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {0027-8424},
number = 14,
volume = 112,
place = {United States},
year = {Tue Apr 07 00:00:00 EDT 2015},
month = {Tue Apr 07 00:00:00 EDT 2015}
}

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Cited by: 36 works
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Novel protein science enabled by total chemical synthesis
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Regulation of Three Virulence Strategies of Mycobacterium tuberculosis: A Success Story
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Mycobacterial Dormancy Systems and Host Responses in Tuberculosis
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