22-26 March 2021
Zoom Conference
Africa/Johannesburg timezone
Biophysics in and for Africa

A Systematic Integration of Empirical and Computational Studies to Biophysically Describe Recombinant Nicotinate Mononucleotide Adenylyltransferase (NaMNAT) From Klebsiella pneumonia

22 Mar 2021, 16:00
20m
Zoom Conference

Zoom Conference

Virtual Event
Oral Presentation Molecular biophysics Molecular biophysics

Speaker

Olamide Jeje

Description

Nicotinate mononucleotide adenylyltransferase (NaMNAT) is an indispensable enzyme in the biosynthesis of pyridine dinucleotides. Given the vital role of NAD+ in controlling key cellular processes, NaMNAT represents an attractive target for the design of novel broad-spectrum antibiotics to treat nosocomial infections associated with MDR Klebsiella Pneumonia. This study aims to characterize the biophysical structure of NaMNAT from K. Pneumonia (KpNaMNAT) using a systematic combination of experimental and computational approaches. Overexpression and purification were carried out using hexa-histidine tags in E. coli expression system and nickel ion-immobilized metal affinity chromatography. Activity studies using NMN substrate showed KpNaMNAT to demonstrate broad pH optima of 6.5-9.5 and preference for Mg2+. Structural characterisation revealed KpNaMNAT as a monomer with predominate α-helices. ATP, NMN, and NAD+ all bind at the same site on KpNaMNAT, but do not induce any significant conformational changes, however, ATP responds to Mg2+ more than the other ligands and the protein response in the presence of Mg2+. The data and insight provided by this novel research would be useful as a molecular basis for further evaluation of the enzymes for the design of structure-based inhibitors with therapeutic potential.

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