title: Brucella melitensis Methionyl-tRNA-Synthetase (MetRS), a Potential Drug Target for Brucellosis.
abstract: We investigated Brucella melitensis methionyl-tRNA-synthetase (BmMetRS) with molecular, structural and phenotypic methods to learn if BmMetRS is a promising target for brucellosis drug development. Recombinant BmMetRS was expressed, purified from wild type Brucella melitensis biovar Abortus 2308 strain ATCC/CRP #DD-156 and screened by a thermal melt assay against a focused library of one hundred previously classified methionyl-tRNA-synthetase inhibitors of the blood stage form of Trypanosoma brucei. Three compounds showed appreciable shift of denaturation temperature and were selected for further studies on inhibition of the recombinant enzyme activity and cell viability against wild type B. melitensis strain 16M. BmMetRS protein complexed with these three inhibitors resolved into three-dimensional crystal structures and was analyzed. All three selected methionyl-tRNA-synthetase compounds inhibit recombinant BmMetRS enzymatic functions in an aminoacylation assay at varying concentrations. Furthermore, growth inhibition of B. melitensis strain 16M by the compounds was shown. Inhibitor-BmMetRS crystal structure models were used to illustrate the molecular basis of the enzyme inhibition. Our current data suggests that BmMetRS is a promising target for brucellosis drug development. However, further studies are needed to optimize lead compound potency, efficacy and safety as well as determine the pharmacokinetics, optimal dosage, and duration for effective treatment.
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Kayode K
authors: http://orcid.org/0000-0002-2007-2170
authors: KK
authors: Ojo
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Ranae M
authors: RM
authors: Ranade
authors: Department of Biochemistry, University of Washington, Seattle, Washington, United States of America.
authors: Zhongsheng
authors: Z
authors: Zhang
authors: Beryllium, Bainbridge Island, Washington, United States of America.
authors: David M
authors: DM
authors: Dranow
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Janette B
authors: JB
authors: Myers
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Ryan
authors: R
authors: Choi
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Steve
authors: S
authors: Nakazawa Hewitt
authors: Beryllium, Bainbridge Island, Washington, United States of America.
authors: Thomas E
authors: TE
authors: Edwards
authors: Beryllium, Bainbridge Island, Washington, United States of America.
authors: Douglas R
authors: DR
authors: Davies
authors: Beryllium, Bainbridge Island, Washington, United States of America.
authors: Donald
authors: D
authors: Lorimer
authors: Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, United States of America.
authors: Stephen M
authors: SM
authors: Boyle
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Lynn K
authors: LK
authors: Barrett
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Frederick S
authors: FS
authors: Buckner
authors: Department of Biochemistry, University of Washington, Seattle, Washington, United States of America.
authors: Erkang
authors: E
authors: Fan
authors: Center for Emerging and Re-emerging Infectious Diseases (CERID), Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, United States of America.
authors: Wesley C
authors: WC
authors: Van Voorhis
mesh_terms: D000595:Amino Acid Sequence; D017347:Brucella melitensis / Q000187:drug effects* / Q000201:enzymology* / Q000254:growth & development; D002006:Brucellosis / Q000188:drug therapy* / Q000382:microbiology; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D020128:Inhibitory Concentration 50; D008718:Methionine-tRNA Ligase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D004791:Enzyme Inhibitors; D008718:Methionine-tRNA Ligase