TY  - JOUR
ID  - pittir19856
UR  - http://d-scholarship-dev.library.pitt.edu/19856/
IS  - 7
A1  - Burnett, JC
A1  - Ruthel, G
A1  - Stegmann, CM
A1  - Panchal, RG
A1  - Nguyen, TL
A1  - Hermone, AR
A1  - Stafford, RG
A1  - Lane, DJ
A1  - Kenny, TA
A1  - McGrath, CF
A1  - Wipf, P
A1  - Stahl, AM
A1  - Schmidt, JJ
A1  - Gussio, R
A1  - Brunger, AT
A1  - Bavari, S
Y1  - 2007/02/16/
N2  - 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 ?M. 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 ?M.
JF  - Journal of Biological Chemistry
VL  - 282
SN  - 0021-9258
TI  - Inhibition of metalloprotease botulinum serotype A from a pseudo-peptide binding mode to a small molecule that is active in primary neurons
SP  - 5004 
AV  - public
EP  -  5014
ER  -