eprintid: 19856 rev_number: 24 userid: 1419 dir: disk0/00/01/98/56 datestamp: 2013-10-09 16:06:59 lastmod: 2021-06-23 22:55:05 status_changed: 2013-10-09 16:06:59 type: article metadata_visibility: show item_issues_count: 0 eprint_status: archive creators_name: Burnett, JC creators_name: Ruthel, G creators_name: Stegmann, CM creators_name: Panchal, RG creators_name: Nguyen, TL creators_name: Hermone, AR creators_name: Stafford, RG creators_name: Lane, DJ creators_name: Kenny, TA creators_name: McGrath, CF creators_name: Wipf, P creators_name: Stahl, AM creators_name: Schmidt, JJ creators_name: Gussio, R creators_name: Brunger, AT creators_name: Bavari, S creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: creators_email: pwipf@pitt.edu creators_email: creators_email: creators_email: creators_email: creators_email: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: creators_id: PWIPF creators_id: creators_id: creators_id: creators_id: creators_id: title: Inhibition of metalloprotease botulinum serotype A from a pseudo-peptide binding mode to a small molecule that is active in primary neurons ispublished: pub divisions: sch_as_chemistry full_text_status: public abstract: An efficient research strategy integrating empirically guided, structure-based modeling and chemoinformatics was used to discover potent small molecule inhibitors of the botulinum neurotoxin serotype A light chain. First, a modeled binding mode for inhibitor 2-mercapto-3-phenylpropionyl-RATKML (K i = 330 nM) was generated, and required the use of a molecular dynamic conformer of the enzyme displaying the reorientation of surface loops bordering the substrate binding cleft. These flexible loops are conformationally variable in x-ray crystal structures, and the model predicted that they were pivotal for providing complementary binding surfaces and solvent shielding for the pseudo-peptide. The docked conformation of 2-mercapto-3-phenylpropionyl- RATKML was then used to refine our pharmacophore for botulinum serotype A light chain inhibition. Data base search queries derived from the pharmacophore were employed to mine small molecule (non-peptidic) inhibitors from the National Cancer Institute's Open Repository. Four of the inhibitors possess Ki values ranging from 3.0 to 10.0 μM. Of these, NSC 240898 is a promising lead for therapeutic development, as it readily enters neurons, exhibits no neuronal toxicity, and elicits dose-dependent protection of synaptosomal-associated protein (of 25 kDa) in a primary culture of embryonic chicken neurons. Isothermal titration calorimetry showed that the interaction between NSC 240898 and the botulinum A light chain is largely entropy-driven, and occurs with a 1:1 stoichiometry and a dissociation constant of 4.6 μM. date: 2007-02-16 date_type: published publication: Journal of Biological Chemistry volume: 282 number: 7 pagerange: 5004 - 5014 refereed: TRUE issn: 0021-9258 id_number: 10.1074/jbc.M608166200 pmid: 17092934 mesh_headings: Animals mesh_headings: Botulinum Toxins, Type A--chemistry mesh_headings: Botulinum Toxins, Type A--metabolism mesh_headings: Botulism--drug therapy mesh_headings: Botulism--enzymology mesh_headings: Cells, Cultured mesh_headings: Chick Embryo mesh_headings: Metalloproteases--chemistry mesh_headings: Metalloproteases--metabolism mesh_headings: Models, Molecular mesh_headings: Neurons--chemistry mesh_headings: Neurons--enzymology mesh_headings: Protease Inhibitors--chemistry mesh_headings: Protease Inhibitors--metabolism mesh_headings: Protease Inhibitors--therapeutic use chemical_names: Protease Inhibitors chemical_names: Metalloproteases chemical_names: Botulinum Toxins, Type A citation: Burnett, JC and Ruthel, G and Stegmann, CM and Panchal, RG and Nguyen, TL and Hermone, AR and Stafford, RG and Lane, DJ and Kenny, TA and McGrath, CF and Wipf, P and Stahl, AM and Schmidt, JJ and Gussio, R and Brunger, AT and Bavari, S (2007) Inhibition of metalloprotease botulinum serotype A from a pseudo-peptide binding mode to a small molecule that is active in primary neurons. Journal of Biological Chemistry, 282 (7). 5004 - 5014. ISSN 0021-9258 document_url: http://d-scholarship-dev.library.pitt.edu/19856/1/licence.txt