Actions for Structural and Biophysical Characterization of the <i>Mycobacterium tuberculosis<

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Structural and Biophysical Characterization of the <i>Mycobacterium tuberculosis</i> Protein Rv0577, a Protein Associated with Neutral Red Staining of Virulent Tuberculosis Strains and Homologue of the <i>Streptomyces coelicolor</i> Protein KbpA [Structural and Biophysical Characterization of Rv0577, a Protein Associated with Neutral Red Staining of Virulent Strains of <i>Mycobacterium tuberculosis</i> and homolog of the <i>Streptomyces coelicolor</i> protein KbpA] [electronic resource].

Published
Bethesda, Md. : National Institutes of Health (U.S.), 2017.
Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy
Physical Description
pages 4,015-4,027 : digital, PDF file
Additional Creators
Los Alamos National Laboratory, National Institutes of Health (U.S.), United States. Department of Energy, and United States. Department of Energy. Office of Scientific and Technical Information
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Summary
Mycobacterium tuberculosis protein Rv0577 is a prominent antigen in tuberculosis patients, the component responsible for neutral red staining of virulent strains of M. tuberculosis, a putative component in a methylglyoxal detoxification pathway, and an agonist of toll-like receptor 2. It also has an amino acid sequence that is 36% identical to that of Streptomyces coelicolor AfsK-binding protein A (KbpA), a component in the complex secondary metabolite pathways in the Streptomyces genus. To gain insight into the biological function of Rv0577 and the family of KpbA kinase regulators, the crystal structure for Rv0577 was determined to a resolution of 1.75 Å, binding properties with neutral red and deoxyadenosine were surveyed, backbone dynamics were measured, and thermal stability was assayed by circular dichroism spectroscopy. The protein is composed of four approximate repeats with a βαβββ topology arranged radially in consecutive pairs to form two continuous eight-strand β-sheets capped on both ends with an α-helix. The two β-sheets intersect in the center at roughly a right angle and form two asymmetric deep “saddles” that may serve to bind ligands. Nuclear magnetic resonance chemical shift perturbation experiments show that neutral red and deoxyadenosine bind to Rv0577. Binding to deoxyadenosine is weaker with an estimated dissociation constants of 4.1 ± 0.3 mM for saddle 1. Heteronuclear steady-state {1H}–15N nuclear Overhauser effect, T1, and T2 values were generally uniform throughout the sequence with only a few modest pockets of differences. As a result, circular dichroism spectroscopy characterization of the thermal stability of Rv0577 indicated irreversible unfolding upon heating with an estimated melting temperature of 56 °C.
Report Numbers
E 1.99:la-ur--17-24508
la-ur--17-24508
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Note
Published through SciTech Connect.
07/10/2017.
"la-ur--17-24508"
Biochemistry 56 30 ISSN 0006-2960 AM
Garry W. Buchko; Nathaniel Echols; E. Megan Flynn; Ho -Leung Ng; Samuel Stephenson; Heung -Bok Kim; Peter J. Myler; Thomas Charles Terwilliger; Tom Alber; Chang -Yub Kim.
Funding Information
AC52-06NA25396
View MARC record | catkey: 24045199