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Title: Reductive methylation to improve crystallization of the putative oxidoreductase Rv0765c from Mycobacterium tuberculosis

Abstract

The M. tuberculosis protein Rv0765c was cloned, expressed, purified and crystallized. In an attempt to improve the quality of the crystals of Rv0765c, the protein was modified by reductive methylation. The methylated protein crystallized in a new crystal form with profoundly improved diffraction properties. Rv0765c from Mycobacterium tuberculosis was cloned and heterologously expressed in Escherichia coli. It was purified using affinity and size-exclusion chromatographic techniques and crystallized. The native protein crystallized in a hexagonal crystal form which diffracted to 7 Å resolution. In an attempt to improve the quality of the Rv0765c crystals, the protein was modified by reductive methylation using dimethylaminoborane and formaldehyde. The modified protein crystallized under different conditions in a tetragonal crystal form, from which diffraction data could be collected to a resolution of 3.2 Å. In both crystal forms of Rv0765c, the asymmetric unit contained two copies of the protein molecule.

Authors:
 [1]; ;  [2]; ;  [1]
  1. EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, D-22603 Hamburg (Germany)
  2. Max-Planck-Institute for Infection Biology, Schumannstrasse 21/22, D-10117 Berlin (Germany)
Publication Date:
OSTI Identifier:
22360339
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 63; Journal Issue: Pt 6; Other Information: PMCID: PMC2335070; PMID: 17554174; PUBLISHER-ID: bo5020; OAI: oai:pubmedcentral.nih.gov:2335070; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; AFFINITY; CRYSTALLIZATION; CRYSTALS; DIFFRACTION; ESCHERICHIA COLI; FORMALDEHYDE; MOLECULES; PROTEINS; RESOLUTION

Citation Formats

Rauert, Wilko, Eddine, Ali Nasser, Kaufmann, Stefan H. E., Weiss, Manfred S., and Janowski, Robert, E-mail: robert@embl-hamburg.de. Reductive methylation to improve crystallization of the putative oxidoreductase Rv0765c from Mycobacterium tuberculosis. United Kingdom: N. p., 2007. Web. doi:10.1107/S1744309107022506.
Rauert, Wilko, Eddine, Ali Nasser, Kaufmann, Stefan H. E., Weiss, Manfred S., & Janowski, Robert, E-mail: robert@embl-hamburg.de. Reductive methylation to improve crystallization of the putative oxidoreductase Rv0765c from Mycobacterium tuberculosis. United Kingdom. doi:10.1107/S1744309107022506.
Rauert, Wilko, Eddine, Ali Nasser, Kaufmann, Stefan H. E., Weiss, Manfred S., and Janowski, Robert, E-mail: robert@embl-hamburg.de. Fri . "Reductive methylation to improve crystallization of the putative oxidoreductase Rv0765c from Mycobacterium tuberculosis". United Kingdom. doi:10.1107/S1744309107022506.
@article{osti_22360339,
title = {Reductive methylation to improve crystallization of the putative oxidoreductase Rv0765c from Mycobacterium tuberculosis},
author = {Rauert, Wilko and Eddine, Ali Nasser and Kaufmann, Stefan H. E. and Weiss, Manfred S. and Janowski, Robert, E-mail: robert@embl-hamburg.de},
abstractNote = {The M. tuberculosis protein Rv0765c was cloned, expressed, purified and crystallized. In an attempt to improve the quality of the crystals of Rv0765c, the protein was modified by reductive methylation. The methylated protein crystallized in a new crystal form with profoundly improved diffraction properties. Rv0765c from Mycobacterium tuberculosis was cloned and heterologously expressed in Escherichia coli. It was purified using affinity and size-exclusion chromatographic techniques and crystallized. The native protein crystallized in a hexagonal crystal form which diffracted to 7 Å resolution. In an attempt to improve the quality of the Rv0765c crystals, the protein was modified by reductive methylation using dimethylaminoborane and formaldehyde. The modified protein crystallized under different conditions in a tetragonal crystal form, from which diffraction data could be collected to a resolution of 3.2 Å. In both crystal forms of Rv0765c, the asymmetric unit contained two copies of the protein molecule.},
doi = {10.1107/S1744309107022506},
journal = {Acta Crystallographica. Section F},
number = Pt 6,
volume = 63,
place = {United Kingdom},
year = {Fri Jun 01 00:00:00 EDT 2007},
month = {Fri Jun 01 00:00:00 EDT 2007}
}