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Inhibition of metalloprotease botulinum serotype A from a pseudo-peptide binding mode to a small molecule that is active in primary neurons

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

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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.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Burnett, JC
Ruthel, G
Stegmann, CM
Panchal, RG
Nguyen, TL
Hermone, AR
Stafford, RG
Lane, DJ
Kenny, TA
McGrath, CF
Wipf, Ppwipf@pitt.eduPWIPF
Stahl, AM
Schmidt, JJ
Gussio, R
Brunger, AT
Bavari, S
Date: 16 February 2007
Date Type: Publication
Journal or Publication Title: Journal of Biological Chemistry
Volume: 282
Number: 7
Page Range: 5004 - 5014
DOI or Unique Handle: 10.1074/jbc.m608166200
Schools and Programs: Dietrich School of Arts and Sciences > Chemistry
Refereed: Yes
ISSN: 0021-9258
MeSH Headings: Animals; Botulinum Toxins, Type A--chemistry; Botulinum Toxins, Type A--metabolism; Botulism--drug therapy; Botulism--enzymology; Cells, Cultured; Chick Embryo; Metalloproteases--chemistry; Metalloproteases--metabolism; Models, Molecular; Neurons--chemistry; Neurons--enzymology; Protease Inhibitors--chemistry; Protease Inhibitors--metabolism; Protease Inhibitors--therapeutic use
PubMed ID: 17092934
Date Deposited: 09 Oct 2013 16:06
Last Modified: 26 Sep 2022 16:29


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