Structural and Regulatory Elements of HCV NS5B Polymerase – β-Loop and C-Terminal Tail – Are Required for Activity of Allosteric Thumb Site II Inhibitors
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{"title"=>"Structural and regulatory elements of HCV NS5B polymerase - β-loop and C-terminal tail - Are required for activity of allosteric thumb site II inhibitors", "type"=>"journal", "authors"=>[{"first_name"=>"Sarah E.", "last_name"=>"Boyce", "scopus_author_id"=>"12787101500"}, {"first_name"=>"Neeraj", "last_name"=>"Tirunagari", "scopus_author_id"=>"8585974500"}, {"first_name"=>"Anita", "last_name"=>"Niedziela-Majka", "scopus_author_id"=>"6603324125"}, {"first_name"=>"Jason", "last_name"=>"Perry", "scopus_author_id"=>"7401518185"}, {"first_name"=>"Melanie", "last_name"=>"Wong", "scopus_author_id"=>"55614094100"}, {"first_name"=>"Elaine", "last_name"=>"Kan", "scopus_author_id"=>"7006842409"}, {"first_name"=>"Leanna", "last_name"=>"Lagpacan", "scopus_author_id"=>"10838862800"}, {"first_name"=>"Ona", "last_name"=>"Barauskas", "scopus_author_id"=>"55197267000"}, {"first_name"=>"Magdeleine", "last_name"=>"Hung", "scopus_author_id"=>"15825202000"}, {"first_name"=>"Martijn", "last_name"=>"Fenaux", "scopus_author_id"=>"56156576900"}, {"first_name"=>"Todd", "last_name"=>"Appleby", "scopus_author_id"=>"6602872227"}, {"first_name"=>"William J.", "last_name"=>"Watkins", "scopus_author_id"=>"7203059472"}, {"first_name"=>"Uli", "last_name"=>"Schmitz", "scopus_author_id"=>"7006749826"}, {"first_name"=>"Roman", "last_name"=>"Sakowicz", "scopus_author_id"=>"6602130293"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203", "pmid"=>"24416288", "doi"=>"10.1371/journal.pone.0084808", "pui"=>"372763062", "issn"=>"19326203", "sgr"=>"84897488758", "scopus"=>"2-s2.0-84897488758"}, "id"=>"02e9c633-417b-3f1e-9362-759234e5f499", "abstract"=>"Elucidation of the mechanism of action of the HCV NS5B polymerase thumb site II inhibitors has presented a challenge. Current opinion holds that these allosteric inhibitors stabilize the closed, inactive enzyme conformation, but how this inhibition is accomplished mechanistically is not well understood. Here, using a panel of NS5B proteins with mutations in key regulatory motifs of NS5B--the C-terminal tail and β-loop--in conjunction with a diverse set of NS5B allosteric inhibitors, we show that thumb site II inhibitors possess a distinct mechanism of action. A combination of enzyme activity studies and direct binding assays reveals that these inhibitors require both regulatory elements to maintain the polymerase inhibitory activity. Removal of either element has little impact on the binding affinity of thumb site II inhibitors, but significantly reduces their potency. NS5B in complex with a thumb site II inhibitor displays a characteristic melting profile that suggests stabilization not only of the thumb domain but also the whole polymerase. Successive truncations of the C-terminal tail and/or removal of the β-loop lead to progressive destabilization of the protein. Furthermore, the thermal unfolding transitions characteristic for thumb site II inhibitor-NS5B complex are absent in the inhibitor-bound constructs in which interactions between C-terminal tail and β-loop are abolished, pointing to the pivotal role of both regulatory elements in communication between domains. Taken together, a comprehensive picture of inhibition by compounds binding to thumb site II emerges: inhibitor binding provides stabilization of the entire polymerase in an inactive, closed conformation, propagated via coupled interactions between the C-terminal tail and β-loop.", "link"=>"http://www.mendeley.com/research/structural-regulatory-elements-hcv-ns5b-polymerase-%CE%B2loop-cterminal-tail-required-activity-allosteric", "reader_count"=>16, "reader_count_by_academic_status"=>{"Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>3}, "reader_count_by_user_role"=>{"Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>3}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>7, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Chemistry"=>4, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Chemistry"=>{"Chemistry"=>4}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"Saudi Arabia"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1344403"], "description"=>"<p><sup>a</sup> Sensorgrams for the binding of NNIs to Δ21, Δ55 and Δ21-Δ8 along with a table of equilibrium and kinetic parameters of interaction are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084808#pone.0084808.s002\" target=\"_blank\">Figure S2</a> and Table S1 in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084808#pone.0084808.s006\" target=\"_blank\">File S1</a>, respectively.</p><p><sup>b</sup> Fold shift in K<sub>D</sub> for association of NNIs to Δ55 and Δ21-Δ8 was calculated relative to the K<sub>D</sub> determined for binding towards Δ21.</p><p><sup>c</sup> Where K<sub>D</sub> values could not be obtained either due to complex binding kinetics (Palm Site II inhibitor binding to Δ55 and Δ21-Δ8) or super-stoichiometric binding (Palm Site I inhibitor binding to Δ21-Δ8), numerical values have been replaced by n/a (not applicable).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "dissociation", "constants", "binding", "nnis", "fold-shifts"], "article_id"=>898797, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.t001", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Equilibrium_dissociation_constants_K_D_for_binding_of_NNIs_to_916_21_and_fold_shifts_in_K_D_for_association_to_916_55_and_916_21_916_8_determined_by_SPR_/898797", "title"=>"Equilibrium dissociation constants (K<sub>D</sub>) for binding of NNIs to Δ21 and fold-shifts in K<sub>D</sub> for association to Δ55 and Δ21-Δ8 determined by SPR.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344402"], "description"=>"<p>(<b>A</b>) Apo Δ21 thermal unfolding profile (black line) shown with fit to a non-two-state unfolding model (red) with two transitions: major with T<sub>m</sub>1 (dark blue line) and leading shoulder with T<sub>m</sub>2 (light blue line); (<b>B</b>) Thermal unfolding of Δ21 bound to palm site I shows the same profile as observed for apo Δ21. Melting profile of apo Δ21 bound to palm site II inhibitor is similar to the one observed for Δ21-palm site I complex and apo protein (not shown); (<b>C</b>) Thermal unfolding of Δ21 bound to thumb site I inhibitor with fit to a non-two-state model (red line) with two transitions: major with T<sub>m</sub>1 (dark blue line) and a new transition with T<sub>m</sub>3 (green line); (<b>D</b>) Unfolding profile of Δ21 bound to thumb site II inhibitor GS-9669 shows three transitions. T<sub>m</sub>3 is analogous to the midpoint of second transition observed with thumb site I. (<b>E</b>) and (<b>F</b>) Melting profiles for Δ39 and Δ47 bound to thumb site II inhibitor. (<b>G</b>) Average T<sub>m</sub>3 for the third transition in constructs with C-terminal tail truncations. Value of T<sub>m</sub>3 remains constant with standard deviation better than 0.5°C, but ΔT<sub>m</sub>3 increases with increasing C-terminal truncations. (<b>H</b>) Comparison of changes in ΔT<sub>m</sub>1 and ΔT<sub>m</sub>3 for GS-9669 bound NS5B truncation mutants. Although ΔT<sub>m</sub>3 increases, ΔT<sub>m</sub>1 (st dev 0.1°C) decreases with tail truncation, in agreement with destabilization of polymerase.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "nnis", "ns5b"], "article_id"=>898796, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g006", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Binding_of_thumb_domain_NNIs_to_NS5B_results_in_appearance_of_a_new_transition_with_T_m_3_/898796", "title"=>"Binding of thumb domain NNIs to NS5B results in appearance of a new transition with T<sub>m</sub>3.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344399"], "description"=>"<p>Front (<b>A</b>) and side view (90° rotation with cutaway through the finger domain) (<b>B</b>) of NS5B bound to primer/template RNA (dark grey). Side view shows how the β-loop wraps around RNA as it exits the polymerase. Front (<b>C</b>) and side (<b>D</b>) view of the overlay of NS5B Δ21 structure (fingers-pink, palm-light blue, thumb-light green, PDB entry 1C2P) with the model. Overlay shows that the transition from closed inactive polymerase to elongating enzyme requires only small expansion of the finger (red), palm (blue) and thumb (green) domains and major displacement of the β-loop (yellow) and C-terminal tail (magenta). Black circle in (C) denotes the position of double stranded RNA in the model.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "hcv", "ns5b", "bound", "duplex", "rna", "elongation"], "article_id"=>898793, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g003", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Model_of_HCV_NS5B_bound_to_duplex_RNA_in_elongation_mode_/898793", "title"=>"Model of HCV NS5B bound to duplex RNA in elongation mode.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344397"], "description"=>"<p>Structures of HCV NS5B active site nucleotide inhibitor and NNIs targeting four allosteric sites used in this study.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "hcv", "ns5b", "nucleotide", "inhibitor", "nnis", "targeting", "allosteric", "sites"], "article_id"=>898791, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g002", "stats"=>{"downloads"=>0, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structures_of_HCV_NS5B_active_site_nucleotide_inhibitor_and_NNIs_targeting_four_allosteric_sites_used_in_this_study_/898791", "title"=>"Structures of HCV NS5B active site nucleotide inhibitor and NNIs targeting four allosteric sites used in this study.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344406", "https://ndownloader.figshare.com/files/1344407", "https://ndownloader.figshare.com/files/1344408", "https://ndownloader.figshare.com/files/1344409", "https://ndownloader.figshare.com/files/1344410", "https://ndownloader.figshare.com/files/1344411"], "description"=>"<div><p>Elucidation of the mechanism of action of the HCV NS5B polymerase thumb site II inhibitors has presented a challenge. Current opinion holds that these allosteric inhibitors stabilize the closed, inactive enzyme conformation, but how this inhibition is accomplished mechanistically is not well understood. Here, using a panel of NS5B proteins with mutations in key regulatory motifs of NS5B – the C-terminal tail and β-loop – in conjunction with a diverse set of NS5B allosteric inhibitors, we show that thumb site II inhibitors possess a distinct mechanism of action. A combination of enzyme activity studies and direct binding assays reveals that these inhibitors require both regulatory elements to maintain the polymerase inhibitory activity. Removal of either element has little impact on the binding affinity of thumb site II inhibitors, but significantly reduces their potency. NS5B in complex with a thumb site II inhibitor displays a characteristic melting profile that suggests stabilization not only of the thumb domain but also the whole polymerase. Successive truncations of the C-terminal tail and/or removal of the β-loop lead to progressive destabilization of the protein. Furthermore, the thermal unfolding transitions characteristic for thumb site II inhibitor – NS5B complex are absent in the inhibitor – bound constructs in which interactions between C-terminal tail and β-loop are abolished, pointing to the pivotal role of both regulatory elements in communication between domains. Taken together, a comprehensive picture of inhibition by compounds binding to thumb site II emerges: inhibitor binding provides stabilization of the entire polymerase in an inactive, closed conformation, propagated via coupled interactions between the C-terminal tail and β-loop.</p></div>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "elements", "hcv", "ns5b", "polymerase", "c-terminal", "allosteric", "ii"], "article_id"=>898800, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0084808.s001", "https://dx.doi.org/10.1371/journal.pone.0084808.s002", "https://dx.doi.org/10.1371/journal.pone.0084808.s003", "https://dx.doi.org/10.1371/journal.pone.0084808.s004", "https://dx.doi.org/10.1371/journal.pone.0084808.s005", "https://dx.doi.org/10.1371/journal.pone.0084808.s006"], "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structural_and_Regulatory_Elements_of_HCV_NS5B_Polymerase_8211_946_Loop_and_C_Terminal_Tail_8211_Are_Required_for_Activity_of_Allosteric_Thumb_Site_II_Inhibitors_/898800", "title"=>"Structural and Regulatory Elements of HCV NS5B Polymerase – β-Loop and C-Terminal Tail – Are Required for Activity of Allosteric Thumb Site II Inhibitors", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344401"], "description"=>"<p>(<b>A</b>) Representative DSF melting profile for apo NS5B mutant constructs at 5 µM protein concentration are shown. Trend in stability is: Δ21 > Δ39 >> Δ21–AAA ≈ Δ47 > Δ55 > Δ21-Δ8. (<b>B</b>) DSF average T<sub>m</sub> and T<sub>m</sub> shifts relative to apo Δ21 (ΔT<sub>m</sub>) for mutant constructs. (<b>C</b>) Direct binding of NNIs to NS5B constructs determined by upshift of NS5B T<sub>m</sub> in the presence of 25 µM concentration of thumb site II inhibitor. Values represent an average of two or more independent experiments (each run in triplicate) with standard deviations within 0.5 to 1.0°C, unless mentioned otherwise. Heatmap is colored according to the magnitude of ΔT<sub>m</sub> elicited by inhibitor binding. Thumb site II inhibitors show >2°C T<sub>m</sub> upshift for all constructs in agreement with SPR data obtained for selected constructs.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "mutations", "motifs", "ns5b", "inhibitor"], "article_id"=>898795, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g005", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_mutations_in_structural_motifs_C_terminus_946_loop_of_NS5B_on_protein_stability_and_inhibitor_binding_/898795", "title"=>"Effect of mutations in structural motifs (C-terminus, β-loop) of NS5B on protein stability and inhibitor binding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344400"], "description"=>"<p>Numbers represent mean and standard deviation of IC<sub>50</sub> values determined for each inhibitor against the set of NS5B constructs. Heatmap is colored according to IC<sub>50</sub> fold shift relative to Δ21 to show changes in inhibition profile for given NNI across NS5B mutant constructs. Thumb site II inhibitors begin to show significant loss of inhibitory potency as interactions between β-loop and C-terminal residues are disrupted by mutations in the interface (Δ21-AAA, truncations past Δ39 and Δ21-Δ39 mutations). Palm site I inhibitor is affected as well, which is explained by disruption of the inhibitor's interaction with the β-loop and C-terminal residues. Inhibition by thumb site I remains for the most part unaffected by NS5B mutations whereas inhibition by palm site II NNI is reduced due to decrease in binding to NS5B with truncated β-loop and/or C-terminal.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "inhibition", "rdrp", "ns5b", "nuc"], "article_id"=>898794, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g004", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IC_50_values_for_inhibition_of_RdRp_activity_of_NS5B_by_Nuc_3_8242_d_8242_CTP_and_NNIs_/898794", "title"=>"IC<sub>50</sub> values for inhibition of RdRp activity of NS5B by Nuc (3′-d′CTP) and NNIs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1344395"], "description"=>"<p>(<b>A</b>) Thumb site I and thumb site II are located on the thumb domain (green); palm site I and palm site II are at the interface of the three domains, thumb, palm (blue) and fingers (red). GS-9669 inhibitor bound in the thumb site II pocket is shown in stick representation (grey, description of crystal structure of NS5B bound to thumb site II inhibitor GS-9669 will be published elsewhere). The active site is indicated by the cyan circle. The other main structural features shown are the C-terminal tail residues (magenta) which contact the β-loop (yellow). (<b>B</b>) 2D representation of domain structure of polymerase and C-terminal truncation sites Δ21, Δ39, Δ47, Δ55, as well as the β-loop deletion mutant Δ21-Δ8 (deleted residues shown in yellow) and LWF triple A mutant F550A/W551A/L553A. Δ55 is a tag free construct and all others contain C6-His. (<b>C</b>) Location of the mutations relative to the tertiary protein structure. (<b>D</b>) Close-up view of interface between LWF motif (magenta, stick representation) and β-loop (yellow) which is dominated by hydrophobic contacts on the surface of the protein.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme structure", "proteins", "Protein chemistry", "Protein interactions", "protein structure", "Recombinant proteins", "drug discovery", "Macromolecular assemblies", "Small molecules", "biophysics", "Computational biology", "Macromolecular structure analysis", "medicinal chemistry", "ns5b", "polymerase", "nonnucleoside", "inhibitors", "binding", "sites", "constructs"], "article_id"=>898789, "categories"=>["Biological Sciences", "Chemistry"], "users"=>["Sarah E. Boyce", "Neeraj Tirunagari", "Anita Niedziela-Majka", "Jason Perry", "Melanie Wong", "Elaine Kan", "Leanna Lagpacan", "Ona Barauskas", "Magdeleine Hung", "Martijn Fenaux", "Todd Appleby", "William J. Watkins", "Uli Schmitz", "Roman Sakowicz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0084808.g001", "stats"=>{"downloads"=>2, "page_views"=>128, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HCV_NS5B_polymerase_nonnucleoside_inhibitors_binding_sites_and_NS5B_constructs_used_in_studies_/898789", "title"=>"HCV NS5B polymerase nonnucleoside inhibitors binding sites and NS5B constructs used in studies.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-09 03:43:47"}

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Relative Metric

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