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Title: Preparation of the Crystal Complex of Phosphopantetheine Adenylyltransferase from Mycobacterium tuberculosis with Coenzyme A and Investigation of Its Three-Dimensional Structure at 2.1-A Resolution

Abstract

Recombinant phosphopantetheine adenylyltransferase from Mycobacterium tuberculosis (PPAT Mt), which was produced by a high-producing strain and purified to 99%, was used for the crystal growth of the complex of the enzyme with coenzyme A (CoA). Crystals suitable for X-ray diffraction study were obtained by cocrystallization. The crystals belong to sp. gr. R32 and have the unit-cell parameters a = b = 98.840 A, c = 112.880 A, {alpha} = {beta} = 90.00{sup o}, and {gamma} = 120.00{sup o}. The three-dimensional structure of the complex was determined based on X-ray diffraction data collected from the crystals to 2.1 A resolution and refined to Rf = 22.7% and Rfree = 25.93%. Active-site bound coenzyme A was found, and its nearest environment was described. The conformational changes of the enzyme due to ligand binding were revealed. The binding of CoA by tuberculosis phosphopantetheine adenylyltransferase was characterized by comparing the structures of the title complex to a similar complex of PPAT from E. coli (PPAT Ec).

Authors:
;  [1]; ;  [2];  [1]
  1. Russian Academy of Sciences, Shubnikov Institute of Crystallography (Russian Federation)
  2. Russian Academy of Sciences, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation)
Publication Date:
OSTI Identifier:
22054064
Resource Type:
Journal Article
Resource Relation:
Journal Name: Crystallography Reports; Journal Volume: 55; Journal Issue: 6; Other Information: Copyright (c) 2010 Pleiades Publishing, Ltd.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; COENZYMES; COMPLEXES; CONFORMATIONAL CHANGES; CRYSTAL GROWTH; CRYSTALS; ENZYMES; MYCOBACTERIUM TUBERCULOSIS; RESOLUTION; TRIGONAL LATTICES; X-RAY DIFFRACTION

Citation Formats

Timofeev, V. I., Smirnova, E. A., E-mail: inna@ns.crys.ras.ru, Chupova, L. A., Esipov, R. S., and Kuranova, I. P. Preparation of the Crystal Complex of Phosphopantetheine Adenylyltransferase from Mycobacterium tuberculosis with Coenzyme A and Investigation of Its Three-Dimensional Structure at 2.1-A Resolution. United States: N. p., 2010. Web. doi:10.1134/S1063774510060234.
Timofeev, V. I., Smirnova, E. A., E-mail: inna@ns.crys.ras.ru, Chupova, L. A., Esipov, R. S., & Kuranova, I. P. Preparation of the Crystal Complex of Phosphopantetheine Adenylyltransferase from Mycobacterium tuberculosis with Coenzyme A and Investigation of Its Three-Dimensional Structure at 2.1-A Resolution. United States. doi:10.1134/S1063774510060234.
Timofeev, V. I., Smirnova, E. A., E-mail: inna@ns.crys.ras.ru, Chupova, L. A., Esipov, R. S., and Kuranova, I. P. Mon . "Preparation of the Crystal Complex of Phosphopantetheine Adenylyltransferase from Mycobacterium tuberculosis with Coenzyme A and Investigation of Its Three-Dimensional Structure at 2.1-A Resolution". United States. doi:10.1134/S1063774510060234.
@article{osti_22054064,
title = {Preparation of the Crystal Complex of Phosphopantetheine Adenylyltransferase from Mycobacterium tuberculosis with Coenzyme A and Investigation of Its Three-Dimensional Structure at 2.1-A Resolution},
author = {Timofeev, V. I. and Smirnova, E. A., E-mail: inna@ns.crys.ras.ru and Chupova, L. A. and Esipov, R. S. and Kuranova, I. P.},
abstractNote = {Recombinant phosphopantetheine adenylyltransferase from Mycobacterium tuberculosis (PPAT Mt), which was produced by a high-producing strain and purified to 99%, was used for the crystal growth of the complex of the enzyme with coenzyme A (CoA). Crystals suitable for X-ray diffraction study were obtained by cocrystallization. The crystals belong to sp. gr. R32 and have the unit-cell parameters a = b = 98.840 A, c = 112.880 A, {alpha} = {beta} = 90.00{sup o}, and {gamma} = 120.00{sup o}. The three-dimensional structure of the complex was determined based on X-ray diffraction data collected from the crystals to 2.1 A resolution and refined to Rf = 22.7% and Rfree = 25.93%. Active-site bound coenzyme A was found, and its nearest environment was described. The conformational changes of the enzyme due to ligand binding were revealed. The binding of CoA by tuberculosis phosphopantetheine adenylyltransferase was characterized by comparing the structures of the title complex to a similar complex of PPAT from E. coli (PPAT Ec).},
doi = {10.1134/S1063774510060234},
journal = {Crystallography Reports},
number = 6,
volume = 55,
place = {United States},
year = {Mon Nov 15 00:00:00 EST 2010},
month = {Mon Nov 15 00:00:00 EST 2010}
}