TY  - JOUR
N1  - The copyright of this article belongs to Nature
ID  - open3066
UR  - https://www.nature.com/articles/s41598-022-16468-w#:~:text=Here%2C%20we%20have%20identified%20small,of%20O%2Dacetyl%2Dhomoserine.
IS  - 1
A1  - Chaudhary, Deepika
A1  - Singh, Avantika
A1  - Marzuki, Mardiana
A1  - Ghosh, Abhirupa
A1  - Kidwai, Saqib
A1  - Gosain, Tannu Priya
A1  - Chawla, Kiran
A1  - Gupta, Sonu Kumar
A1  - Agarwal, Nisheeth
A1  - Saha, Sudipto
A1  - Kumar, Yashwant
A1  - Thakur, Krishan Gopal
A1  - Singhal, Amit
A1  - Singh, Ramandeep
Y1  - 2022/08/13/
N2  - There is an urgent need to validate new drug targets and identify small molecules that possess activity against both drug-resistant and drug-sensitive bacteria. The enzymes belonging to amino acid biosynthesis have been shown to be essential for growth in vitro, in vivo and have not been exploited much for the development of anti-tubercular agents. Here, we have identified small molecule inhibitors targeting homoserine acetyl transferase (HSAT, MetX, Rv3341) from M. tuberculosis. MetX catalyses the first committed step in L-methionine and S-adenosyl methionine biosynthesis resulting in the formation of O-acetyl-homoserine. Using CRISPRi approach, we demonstrate that conditional repression of metX resulted in inhibition of M. tuberculosis growth in vitro. We have determined steady state kinetic parameters for the acetylation of L-homoserine by Rv3341. We show that the recombinant enzyme followed Michaelis?Menten kinetics and utilizes both acetyl-CoA and propionyl-CoA as acyl-donors. High-throughput screening of a 2443 compound library resulted in identification of small molecule inhibitors against MetX enzyme from M. tuberculosis. The identified lead compounds inhibited Rv3341 enzymatic activity in a dose dependent manner and were also active against HSAT homolog from S. aureus. Molecular docking of the identified primary hits predicted residues that are essential for their binding in HSAT homologs from M. tuberculosis and S. aureus. ThermoFluor assay demonstrated direct binding of the identified primary hits with HSAT proteins. Few of the identified small molecules were able to inhibit growth of M. tuberculosis and S. aureus in liquid cultures. Taken together, our findings validated HSAT as an attractive target for development of new broad-spectrum anti-bacterial agents that should be effective against drug-resistant bacteria.
PB  - Nature 
JF  - SCIENTIFIC REPORTS
VL  - 12
TI  - Identification of small molecules targeting homoserine acetyl transferase from Mycobacterium tuberculosis and Staphylococcus aureus
ER  -