The Dephospho-CoA Kinase of Mycobacterium tuberculosis: Molecular Mechanism and Function.
TL;DR
The found that the S15A, R145A or N175A mutants showed markedly reduced activity, whereas the K14A mutation completely abolished activity, and deletion of residues 27-101 (Δ27-101) led to a significant loss of function.
Abstract
Tuberculosis (TB) continues to represent a significant global health danger, with the bacillus Mycobacterium tuberculosis (Mtb) identified as its causative agent. An essential metabolic helper, Coenzyme A (CoA) engages in multiple core cellular processes that are pivotal for both Mtb survival and its ability to cause disease. The terminal step of CoA biosynthesis is catalyzed by dephospho-CoA kinase CoaE, which phosphorylates dephospho-CoA to generate the active cofactor. However, the molecular mechanisms regulating Mtb CoaE catalytic activity remain poorly understood. Here, AlphaFold 3 was used to predict CoaE model. The Ramachandran plot and ProSA were used to validate the model. Subsequently, we performed molecular docking, followed by site-directed mutagenesis. We found that the S15A, R145A or N175A mutants showed markedly reduced activity, whereas the K14A mutation completely abolished activity. Additionally, deletion of residues 27-101 (Δ27-101) led to a significant loss of function. Our findings offer new insights into the structure and mechanism of Mtb CoaE, a promising target for designing selective antituberculosis agents.