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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Binding studies of a putative C. pseudotuberculosis target protein from Vitamin B-12 Metabolism

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Peinado, Rafaela dos S. [1] ; Olivier, Danilo S. [1] ; Eberle, Raphael J. [1] ; de Moraes, Fabio R. [1] ; Amaral, Marcos S. [2] ; Arni, Raghuvir K. [1] ; Coronado, Monika A. [1]
Total Authors: 7
[1] Univ Estadual Paulista UNESP, Multiuser Ctr Biomol Innovat, Dept Phys, Inst Biociencias Letras & Ciencias Exatas Ibilce, BR-15054000 Sao Jose Do Rio Preto, SP - Brazil
[2] Univ Fed Mato Grosso do Sul, Inst Phys, BR-79090700 Campo Grande, MS - Brazil
Total Affiliations: 2
Document type: Journal article
Source: SCIENTIFIC REPORTS; v. 9, APR 23 2019.
Web of Science Citations: 0

Vitamin B-12 acts as a cofactor for various metabolic reactions important in living organisms. The Vitamin B-12 biosynthesis is restricted to prokaryotes, which means, all eukaryotic organisms must acquire this molecule through diet. This study presents the investigation of Vitamin B-12 metabolism and the characterization of precorrin-4 C(11)-methyltransferase (CobM), an enzyme involved in the biosynthesis of Vitamin B-12 in Corynebacterium pseudotuberculosis. The analysis of the C. pseudotuberculosis genome identified two Vitamin B-12-dependent pathways, which can be strongly affected by a disrupted vitamin metabolism. Molecular dynamics, circular dichroism, and NMR-STD experiments identified regions in CobM that undergo conformational changes after s-adenosyl-L-methionine binding to promote the interaction of precorrin-4, a Vitamin B(12 )precursor. The binding of s-adenosyl-L-methionine was examined along with the competitive binding of adenine, dATP, and suramin. Based on fluorescence spectroscopy experiments the dissociation constant for the four ligands and the target protein could be determined; SAM (1.4 +/- 0.7 mu M), adenine (17.8 +/- 1.5 mu M), dATP (15.8 +/- 2.0 mu M), and Suramin (6.3 +/- 1.1 mu M). The results provide rich information for future investigations of potential drug targets within the C. pseudotuberculosis's Vitamin B12 metabolism and related pathways to reduce the pathogen's virulence in its hosts. (AU)

FAPESP's process: 09/53989-4 - Acquisition of a nuclear magnetic resonance spectrometer for studies of biomolecules
Grantee:Raghuvir Krishnaswamy Arni
Support Opportunities: Multi-user Equipment Program
FAPESP's process: 15/18868-2 - Multi-user equipment acquisition for molecular and structural biology
Grantee:Raghuvir Krishnaswamy Arni
Support Opportunities: Multi-user Equipment Program
FAPESP's process: 16/08104-8 - Structural and functional aspects of two DNA binding proteins encoded by Corynebacterium pseudotuberculosis
Grantee:Raphael Josef Eberle
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 15/13765-0 - Structural Studies and Characterization of Proteins by X-ray Crystallography and Nuclear Magnetic Resonance. Structural investigations and biophysics of molecular mechanisms of functional proteins.
Grantee:Raghuvir Krishnaswamy Arni
Support Opportunities: Regular Research Grants
FAPESP's process: 16/12904-0 - Mechanism and Molecular Interactions of Bioactive molecules with NS3 protease from Zika virus.
Grantee:Monika Aparecida Coronado
Support Opportunities: Scholarships in Brazil - Post-Doctoral