Biochemical and Structural Characterization of Mycobacterial Aspartyl-tRNA Synthetase AspS, a Promising TB Drug Target
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{"title"=>"Biochemical and structural characterization of mycobacterial aspartyl-tRNA synthetase AspS, a promising TB drug target", "type"=>"journal", "authors"=>[{"first_name"=>"Sudagar S.", "last_name"=>"Gurcha", "scopus_author_id"=>"6602113921"}, {"first_name"=>"Veeraraghavan", "last_name"=>"Usha", "scopus_author_id"=>"14064067500"}, {"first_name"=>"Jonathan A.G.", "last_name"=>"Cox", "scopus_author_id"=>"55542146100"}, {"first_name"=>"Klaus", "last_name"=>"Fütterer", "scopus_author_id"=>"6701315459"}, {"first_name"=>"Katherine A.", "last_name"=>"Abrahams", "scopus_author_id"=>"55542599000"}, {"first_name"=>"Apoorva", "last_name"=>"Bhatt", "scopus_author_id"=>"16479804000"}, {"first_name"=>"Luke J.", "last_name"=>"Alderwick", "scopus_author_id"=>"8908289900"}, {"first_name"=>"Robert C.", "last_name"=>"Reynolds", "scopus_author_id"=>"7401829883"}, {"first_name"=>"Nicholas J.", "last_name"=>"Loman", "scopus_author_id"=>"6701414703"}, {"first_name"=>"Vijayashankar", "last_name"=>"Nataraj", "scopus_author_id"=>"56440226800"}, {"first_name"=>"Carlos", "last_name"=>"Alemparte", "scopus_author_id"=>"57194286516"}, {"first_name"=>"David", "last_name"=>"Barros", "scopus_author_id"=>"56653725900"}, {"first_name"=>"Adrian J.", "last_name"=>"Lloyd", "scopus_author_id"=>"26643271600"}, {"first_name"=>"Lluis", "last_name"=>"Ballell", "scopus_author_id"=>"6508364138"}, {"first_name"=>"Judith V.", "last_name"=>"Hobrath", "scopus_author_id"=>"14055958700"}, {"first_name"=>"Gurdyal S.", "last_name"=>"Besra", "scopus_author_id"=>"7004651537"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84915803426", "sgr"=>"84915803426", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0113568", "pmid"=>"25409504", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pui"=>"600653148"}, "id"=>"29b91bb0-2569-396b-8be0-1d9c30f5aadf", "abstract"=>"The human pathogen Mycobacterium tuberculosis is the causative agent of pulmonary tuberculosis (TB), a disease with high worldwide mortality rates. Current treatment programs are under significant threat from multi-drug and extensively-drug resistant strains of M. tuberculosis, and it is essential to identify new inhibitors and their targets. We generated spontaneous resistant mutants in Mycobacterium bovis BCG in the presence of 10× the minimum inhibitory concentration (MIC) of compound 1, a previously identified potent inhibitor of mycobacterial growth in culture. Whole genome sequencing of two resistant mutants revealed in one case a single nucleotide polymorphism in the gene aspS at 535GAC>535AAC (D179N), while in the second mutant a single nucleotide polymorphism was identified upstream of the aspS promoter region. We probed whole cell target engagement by overexpressing either M. bovis BCG aspS or Mycobacterium smegmatis aspS, which resulted in a ten-fold and greater than ten-fold increase, respectively, of the MIC against compound 1. To analyse the impact of inhibitor 1 on M. tuberculosis AspS (Mt-AspS) activity we over-expressed, purified and characterised the kinetics of this enzyme using a robust tRNA-independent assay adapted to a high-throughput screening format. Finally, to aid hit-to-lead optimization, the crystal structure of apo M. smegmatis AspS was determined to a resolution of 2.4 Å.", "link"=>"http://www.mendeley.com/research/biochemical-structural-characterization-mycobacterial-aspartyltrna-synthetase-asps-promising-tb-drug", "reader_count"=>32, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>9, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>3, "Student > Bachelor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>9, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>3, "Student > Bachelor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Nursing and Health Professions"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Mathematics"=>1, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Chemistry"=>5, "Computer Science"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Chemistry"=>{"Chemistry"=>5}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Computer Science"=>{"Computer Science"=>1}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>4}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"United States"=>2, "Brazil"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1801466"], "description"=>"<p>Only amino acids forming direct interactions with compound <b>1</b> are shown. Carbon atoms of AspS are colored light green except for carbons of resistance conferring mutant site residues, which are colored purple. Carbons of compound <b>1</b> are shown in orange; all other atoms are colored by atom type (Oxygen red, Nitrogen blue, Sulphur yellow, Chlorine dark green, Hydrogen white). Labels of residues participating in polar interactions are colored brown, labels of all other residues are in black. Polar interactions are indicated with dashed lines and are numbered. Distances between heavy atoms of the atoms groups participating in polar interactions are as follows: <i>1</i>. 3.1 Å; <i>2</i>. 3.8 Å; <i>3</i>. 3.0 Å; <i>4</i>. 3.2 Å; <i>5</i>. 3.1 Å; <i>6</i>. 3.7 Å.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247309, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g009", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Induced_Fit_docked_pose_of_compound_1_at_the_M_smegmatis_AspS_crystal_structure_/1247309", "title"=>"Induced Fit docked pose of compound 1 at the <i>M. smegmatis</i> AspS crystal structure.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801465"], "description"=>"<p>The sites of resistance conferring mutations are indicated by spheres in yellow, and the aspartyl adenylate donor substrate (AMO), positioned according to the superposition with <i>E. coli</i> AspRS (PDB entry 1C0A, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113568#pone.0113568-Eiler1\" target=\"_blank\">[61]</a>) is indicated by spheres in turquoise.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247308, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g008", "stats"=>{"downloads"=>0, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Molecular_surface_of_Ms_AspS_showing_the_inhibitor_binding_pocket_adjacent_to_the_aspartyl_adenylate_binding_site_showing_the_position_of_the_aspartyl_adenylate_donor_substrate_relative_to_the_proposed_pocket_/1247308", "title"=>"Molecular surface of Ms-AspS showing the inhibitor-binding pocket adjacent to the aspartyl adenylate binding site, showing the position of the aspartyl adenylate donor substrate relative to the proposed pocket.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801462"], "description"=>"<p>The reaction mixture was added to increasing amounts of <b>1</b> and <b>2</b>, and the reaction initiated with the substrate PP<i><sub>i</sub></i> and the fluorescence intensity read using a microtitre plate reader. The assays were performed in triplicate and the percentage activity plotted <i>versus</i> the log µM concentration of <b>1</b> and <b>2</b>.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247305, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g005", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dose_response_in_vitro_activity_curves_of_the_effect_of_compound_1_A_and_compound_2_B_on_Mt_AspS_/1247305", "title"=>"Dose response <i>in vitro</i> activity curves of the effect of compound 1 (A) and compound 2 (B) on Mt-AspS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801460"], "description"=>"<p>The overexpression plasmids pMV261, pMV261-BCG aspS and pMV261-Ms aspS were electroporated into M. bovis BCG with kanamycin at 25 µg/ml, and the MIC and resistance against compound <b>1</b> and <b>2</b> was evaluated.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247303, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g003", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Whole_cell_target_based_resistance_using_overexpression_of_aspS_in_M_bovis_BCG_against_compound_1_and_2_/1247303", "title"=>"Whole cell target based resistance using overexpression of aspS in <i>M. bovis</i> BCG against compound 1 and 2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801461"], "description"=>"<p>Michaelis-Menten curves were fitted for <b>A</b>) varying ADPCP as substrate. <b>B</b>) <b>L</b>-Asp as substrate at fixed saturating concentrations of ADPCP and PP<i><sub>i</sub>.</i><b>C</b>) PP<i><sub>i</sub></i> as substrate at fixed concentrations of ADPCP and <b>L</b>-Asp. <b>D</b>) ADPNP as substrate. <b>E</b>) <b>L</b>-Asp as substrate at fixed concentrations of ADPNP and PP<i><sub>i</sub></i>. <b>F</b>) ATP dependence of hexokinase/glucose-6-phosphate dehydrogenase activity. The initial velocity data (dA/min) were plotted against the substrate concentration. Each assay was done in triplicate and expressed as mean ± standard error of mean.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247304, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g004", "stats"=>{"downloads"=>0, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Substrate_dependence_of_Mt_AspS_activity_/1247304", "title"=>"Substrate dependence of Mt-AspS activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801459"], "description"=>"<p>RT-PCR was performed with RT (upper panel) and without RT (lower panel). The RT-PCR cycling conditions were programmed for 25, 30, 40 and 50 cycles, respectively, to get sufficient yields of the 200 bp amplified product and analysed on a 2% agarose gel following staining.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247302, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g002", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_the_expression_levels_of_the_transcript_produced_by_wild_type_M_bovis_BCG_aspS_mutant_1_and_promoter_up_mutant_2_by_RT_PCR_/1247302", "title"=>"Comparison of the expression levels of the transcript produced by wild type <i>M. bovis</i> BCG, <i>aspS</i> mutant 1 and promoter up mutant 2 by RT-PCR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801470"], "description"=>"1<p>Numbers in parentheses refer to the last shell. <sup>2</sup>Ramachandran statistics were determined using Molprobity <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113568#pone.0113568-Davis1\" target=\"_blank\">[62]</a>.</p><p>Statistics of X-ray diffraction data and crystallographic structure refinement.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247313, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.t004", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Statistics_of_X_ray_diffraction_data_and_crystallographic_structure_refinement_/1247313", "title"=>"Statistics of X-ray diffraction data and crystallographic structure refinement.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801468"], "description"=>"a<p>Genomic positions are relative to <i>M. bovis</i> BCG str. Pasteur 1173P2 chromosome (Genbank accession: NC_008769.1). Mutations are reported relative to <i>aspS</i> which is encoded on the reverse strand.</p><p>Single nucleotide polymorphisms detected in <i>M. bovis</i> BCG spontaneous resistant mutants.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247311, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.t002", "stats"=>{"downloads"=>2, "page_views"=>35, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_nucleotide_polymorphisms_detected_in_M_bovis_BCG_spontaneous_resistant_mutants_/1247311", "title"=>"Single nucleotide polymorphisms detected in <i>M. bovis</i> BCG spontaneous resistant mutants.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801469"], "description"=>"<p>Summary of K<i><sub>m</sub></i> determined for the Mt-AspS substrates and the hexokinase/glucose-6-phosphate dehydrogenase substrate.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247312, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.t003", "stats"=>{"downloads"=>3, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Summary_of_K_m_determined_for_the_Mt_AspS_substrates_and_the_hexokinase_glucose_6_phosphate_dehydrogenase_substrate_/1247312", "title"=>"Summary of K<i><sub>m</sub></i> determined for the Mt-AspS substrates and the hexokinase/glucose-6-phosphate dehydrogenase substrate.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801467"], "description"=>"<p>Primers used in this study.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247310, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.t001", "stats"=>{"downloads"=>6, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primers_used_in_this_study_/1247310", "title"=>"Primers used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801464"], "description"=>"<p>Bars in blue, green and orange indicate the N-terminal, catalytic and insertion domains of AspS, respectively. Asterisks indicate the sites of resistance-conferring mutations.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247307, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g007", "stats"=>{"downloads"=>1, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Alignment_of_the_amino_acid_sequences_of_AspS_of_M_smegmatis_Msm_M_tuberculosis_Mtb_and_M_bovis_Mbv_with_that_of_E_coli_AspRS_Eco_/1247307", "title"=>"Alignment of the amino acid sequences of AspS of <i>M. smegmatis</i> (Msm), <i>M. tuberculosis</i> (Mtb) and <i>M. bovis</i> (Mbv) with that of <i>E. coli</i> AspRS (Eco).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801463"], "description"=>"<p><b>A</b>) Dimer of Ms-AspS in ribbon representation (colored by monomer) with resistance-conferring mutation sites indicated in yellow spheres. The binding site of aspartyl adenylate (AMO, spheres in cyan) is derived from the secondary structure-matched superposition with the structure of <i>E. coli</i> AspRS (PDB entry 1C0A, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113568#pone.0113568-Eiler1\" target=\"_blank\">[61]</a>). <b>B</b>) Superposition of the Ms<i>-</i>AspS monomer with the tRNA-bound structure <i>E. coli</i> AspRS (grey ribbon, PDB entry 1C0A). <b>C</b>) Detail of the superposition of Ms-AspS (blue ribbon), PDB entry 4O2D (green ribbon) of the same protein and <i>E. coli</i> AspRS (grey ribbon), focusing on the flexible loop spanning residues 427 to 444. To illustrate the variable conformation of the loop, sticks indicate the spatial positions of Trp444 and Asp436.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247306, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g006", "stats"=>{"downloads"=>1, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structure_of_M_smegmatis_AspS_/1247306", "title"=>"Structure of <i>M. smegmatis</i> AspS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1801458"], "description"=>"<p>Structures of compounds 1 and 2 used in this study.</p>", "links"=>[], "tags"=>["mic", "179N", "Whole Genome Sequencing", "compound", "gac", "cell target engagement", "asps", "Promising TB Drug Target", "bovis BCG aspS", "pathogen Mycobacterium tuberculosis", "nucleotide polymorphism", "Å.", "Current treatment programs", "Mycobacterium smegmatis aspS", "inhibitor", "mutant", "Mycobacterium bovis BCG", "aspS promoter region"], "article_id"=>1247301, "categories"=>["Biological Sciences"], "users"=>["Sudagar S. Gurcha", "Veeraraghavan Usha", "Jonathan A. G. Cox", "Klaus Fütterer", "Katherine A. Abrahams", "Apoorva Bhatt", "Luke J. Alderwick", "Robert C. Reynolds", "Nicholas J. Loman", "VijayaShankar Nataraj", "Carlos Alemparte", "David Barros", "Adrian J. Lloyd", "Lluis Ballell", "Judith V. Hobrath", "Gurdyal S. Besra"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0113568.g001", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structures_of_compounds_1_and_2_used_in_this_study_/1247301", "title"=>"Structures of compounds 1 and 2 used in this study.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-11-19 15:46:54"}

PMC Usage Stats | Further Information

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