?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F34411%2F&rft.title=Identification+of+novel+bacterial+MurA+inhibitors&rft.creator=Shaw%2C+JoAnna+F.&rft.description=Antibiotic+resistance+is+a+persistent+and+serious+public+health+issue+which+causes+many+illnesses+and+deaths+per+year.+In+contrast+to+the+rapid+increase+and+spread+of+drug-resistant+bacteria%2C+antibiotic+development+has+slowed%2C+and+there+is+a+clear+need+to+identify+and+develop+antibiotics+with+new+scaffolds+and+mechanisms+of+action.+In+particular%2C+there+is+a+specific+need+for+novel+antimicrobial+agents+which+are+active+against+both+gram+negative+and+gram+positive+pathogens.+The+bacterial+enzyme+MurA+catalyzes+the+transfer+of+enolpyruvate+from+phosphoenolpyruvate+(PEP)+to+uridine+diphospho-N-acetylglucosamine+(UNAG)%2C+which+is+the+first+committed+step+of+bacterial+cell+wall+biosynthesis.+Currently%2C+the+only+antibiotic+targeted+toward+MurA+is+fosfomycin%2C+which+inhibits+MurA+by+forming+a+covalent+bond+with+MurA%E2%80%99s+active+site+residue%2C+Cys115.+However%2C+MurA+variants+which+lack+the+cysteine+residue+in+the+active+site+(e.g.+M.+tuberculosis+MurA+and+some+vancomycin+resistant+Enterococcus+(VRE)+strains)+are+resistant+to+fosfomycin.%0D%0AThe+goal+of+this+study+was+to+identify+novel+MurA+inhibitors+with+a+different+mechanism+of+action+than+fosfomycin%2C+and+which+are+active+against+a+range+of+gram+negative+and+gram+positive+bacteria.+To+this+end%2C+we+have+developed+and+optimized+an+in+vivo+high-throughput+screening+assay+to+test+recombinant+MurA+enzyme+against+the+TimTec+ApexScreen+library%2C+a+drug+library+with+5%2C040+structurally+diverse+compounds.+The+hits+identified+from+this+screen+were+further+validated+using+different+assays%2C+including+bacterial+growth+curves.+Minimum+Inhibitory+Concentration+(MIC)+values+were+determined+for+the+three+most+promising+hits.+Toxicity+assays+for+these+hits+were+also+conducted+in+two+cell+lines%2C+and+selectivity+indexes+for+each+inhibitor+were+calculated.+Future+directions+include+further+characterizing+the+mechanisms+of+action+and+testing+for+toxicity+in+other+cell+lines+than+HeLa+and+293T%2C+including+primary+cells.&rft.date=2018-06-28&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F34411%2F1%2FShawJoAnna_MSthesis_April2018.pdf&rft.identifier=++Shaw%2C+JoAnna+F.++(2018)+Identification+of+novel+bacterial+MurA+inhibitors.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++