You are here : Home > The lab > Structure and mechanism of Quinolinate Synthase: An enzyme with a [4Fe-4S]2+ cluster and an antibacterial target

Alice Chan

Structure and mechanism of Quinolinate Synthase: An enzyme with a [4Fe-4S]2+ cluster and an antibacterial target

Published on 5 December 2014


Thesis presented December 05, 2014

Abstract :
The Nicotinamide Adenine Dinucleotide (NAD) is a key cofactor essential for cellular metabolism. Synthesized from quinolinic acid (QA) in all living organisms, NAD biosynthesis is different between eukaryotes and prokaryotes​. Indeed, most of eukaryotes produce QA from L-tryptophan, whereas most of prokaryotes and plants synthesize QA by the concerted action of 2 enzymes: L-aspartate oxydase (NadB), an FAD enzyme, which catalyzes L-Aspartate oxidation to form iminoaspartate (IA) while quinolinate synthetase (NadA) allows condensation between IA and Dihydroxyacetone Phosphate (DHAP) to produce QA. Besides this « de novo » pathway, most eukaryotes and some bacteriae have a salvage pathway which allows NAD synthesis from nutrients and metabolites of NAD degradation in order to maintain a correct pool of NAD in the cell. However, some pathogens like Mycobacterium leprae, Helicobacter pylori do not possess this pathway. As a consequence, NadA represents a very attractive target for designing specific antibacterial agents since it does not exist in Human. NadA is the only metalloenzyme of NAD de novo biosynthesis whose molecular mechanism and tridimensional structure with its [4Fe-4S]2+ cluster are unknown. Using substrate and intermediate analogues, we have been able to understand better NadA mechanism, especially [4Fe-4S]2+ cluster role in catalysis. Moreover, we proposed the first in vitro and in vivo inhibitor of NadA: the 4,5 Dithiohydroxyphtalic Acid (DTHPA) which gave us basis to design powerful and specific NadA inhibitors thanks to a structure-activity relationship study. Besides, we resolved the first X-rays structure of NadA under its holoprotein form. Datas we extracted from it helped us greatly to understand NadA mechanism.


Keywords:
Quinolinate synthase, Iron-sulfur, NAD, inhibitors, mechanism, metalloenzyme

Download this thesis (Intranet link).