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Targeting Aspartate Transcarbamoylase in Staphylococcus aureus : A Novel Therapeutic Approach for Sepsis
Author(s) -
Patel Chandni,
Vaishnav Asmita,
Brunzelle Joseph S.,
GuyEvans Hedeel I.,
Edwards Brian FP.,
Evans David R.
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.797.12
Subject(s) - aquifex aeolicus , staphylococcus aureus , aspartate carbamoyltransferase , escherichia coli , sepsis , pyrimidine , pyrimidine metabolism , chemistry , bacteria , antibiotics , pseudomonas aeruginosa , enzyme , microbiology and biotechnology , biochemistry , biology , gene , immunology , genetics , purine
Sepsis is one of the leading causes of death in the United States. More than 1.5 million people get sepsis and 250,000 people die from sepsis each year. Staphylococcus aureus happens to be most common cause of healthcare‐associated infections. Research and development of new antibiotics against S. aureus is a high priority due to its multi‐drug resistance. The levels of pyrimidines in blood are too low to sustain the growth of bacteria, so they must rely on pyrimidine biosynthesis. Previous studies have shown that a defect in several pyrimidine biosynthetic enzymes resulted in 1000‐fold decrease in the titer of bacteria growing in blood. Aspartate transcarbamoylase (ATC) catalyzes the first committed step of the de novo pyrimidine pathway, making it a good drug target. S. aureus ATC containing his‐tag was overexpressed in Escherichia coli and purified using nickel affinity column chromatography. S. aureus ATC has a K m of 2.4 mM for carbamoyl phosphate and 4.5 mM for aspartate, with a maximal velocity of 296 μmol/min/mg. Cross‐linking data suggests that S. aureus ATC is a trimer of 33 kDa and it does not associate with DHO, the subsequent enzyme in the pathway, as seen in Pseudomonas aeruginosa, Aquifex aeolicus, and several other ATC. We have successfully solved the preliminary structure of S. aureus ATC at 2.27Å resolution. Once our preliminary structure has been further refined, it will serve as the basis for the design of novel non‐competitive inhibitors that block domain closure of ATC, an essential step in catalysis. Support or Funding Information American Heart Association Predoctoral Fellowship, Interdisciplinary Biomedical Sciences Fellowship, Wayne State University School of Medicine, Wayne State University Graduate School & Office of VP for Research This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .