skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors

Abstract

Because of their central role in programmed cell death, the caspases are attractive targets for developing new therapeutics against cancer and autoimmunity, myocardial infarction and ischemic damage, and neurodegenerative diseases. We chose to target caspase-3, an executioner caspase, and caspase-8, an initiator caspase, based on the vast amount of information linking their functions to diseases. Through a structure-based drug design approach, a number of novel {beta}-strand peptidomimetic compounds were synthesized. Kinetic studies of caspase-3 and caspase-8 inhibition were carried out with these urazole ring-containing irreversible peptidomimetics and a known irreversible caspase inhibitor, Z-VAD-fmk. Using a stopped-flow fluorescence assay, we were able to determine individual kinetic parameters of caspase-3 and caspase-8 inhibition by these inhibitors. Z-VAD-fmk and the peptidomimetic inhibitors inhibit caspase-3 and caspase-8 via a three-step kinetic mechanism. Inhibition of both caspase-3 and caspase-8 by Z-VAD-fmk and of caspase-3 by the peptidomimetic inhibitors proceeds via two rapid equilibrium steps followed by a relatively fast inactivation step. However, caspase-8 inhibition by the peptidomimetics goes through a rapid equilibrium step, a slow-binding reversible step, and an extremely slow inactivation step. The crystal structures of inhibitor complexes of caspases-3 and -8 validate the design of the inhibitors by illustrating in detail how theymore » mimic peptide substrates. One of the caspase-8 structures also shows binding at a secondary, allosteric site, providing a possible route to the development of noncovalent small molecule modulators of caspase activity.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ;  [1];  [2]
  1. (USC)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1002641
Resource Type:
Journal Article
Journal Name:
BBA- Proteins Proteom.
Additional Journal Information:
Journal Volume: 1804; Journal Issue: (9) ; 09, 2010; Journal ID: ISSN 1570-9639
Country of Publication:
United States
Language:
ENGLISH
Subject:
36 MATERIALS SCIENCE; CRYSTAL STRUCTURE; DEATH; DESIGN; DISEASES; FLUORESCENCE; INACTIVATION; KINETICS; MYOCARDIAL INFARCTION; NEOPLASMS; PEPTIDES; SUBSTRATES; TARGETS

Citation Formats

Wang, Zhigang, Watt, William, Brooks, Nathan A., Harris, Melissa S., Urban, Jan, Boatman, Douglas, McMillan, Michael, Kahn, Michael, Heinrikson, Robert L., Finzel, Barry C., Wittwer, Arthur J., Blinn, James, Kamtekar, Satwik, Tomasselli, Alfredo G., and Pfizer). Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors. United States: N. p., 2010. Web. doi:10.1016/j.bbapap.2010.05.007.
Wang, Zhigang, Watt, William, Brooks, Nathan A., Harris, Melissa S., Urban, Jan, Boatman, Douglas, McMillan, Michael, Kahn, Michael, Heinrikson, Robert L., Finzel, Barry C., Wittwer, Arthur J., Blinn, James, Kamtekar, Satwik, Tomasselli, Alfredo G., & Pfizer). Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors. United States. doi:10.1016/j.bbapap.2010.05.007.
Wang, Zhigang, Watt, William, Brooks, Nathan A., Harris, Melissa S., Urban, Jan, Boatman, Douglas, McMillan, Michael, Kahn, Michael, Heinrikson, Robert L., Finzel, Barry C., Wittwer, Arthur J., Blinn, James, Kamtekar, Satwik, Tomasselli, Alfredo G., and Pfizer). Fri . "Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors". United States. doi:10.1016/j.bbapap.2010.05.007.
@article{osti_1002641,
title = {Kinetic and structural characterization of caspase-3 and caspase-8 inhibition by a novel class of irreversible inhibitors},
author = {Wang, Zhigang and Watt, William and Brooks, Nathan A. and Harris, Melissa S. and Urban, Jan and Boatman, Douglas and McMillan, Michael and Kahn, Michael and Heinrikson, Robert L. and Finzel, Barry C. and Wittwer, Arthur J. and Blinn, James and Kamtekar, Satwik and Tomasselli, Alfredo G. and Pfizer)},
abstractNote = {Because of their central role in programmed cell death, the caspases are attractive targets for developing new therapeutics against cancer and autoimmunity, myocardial infarction and ischemic damage, and neurodegenerative diseases. We chose to target caspase-3, an executioner caspase, and caspase-8, an initiator caspase, based on the vast amount of information linking their functions to diseases. Through a structure-based drug design approach, a number of novel {beta}-strand peptidomimetic compounds were synthesized. Kinetic studies of caspase-3 and caspase-8 inhibition were carried out with these urazole ring-containing irreversible peptidomimetics and a known irreversible caspase inhibitor, Z-VAD-fmk. Using a stopped-flow fluorescence assay, we were able to determine individual kinetic parameters of caspase-3 and caspase-8 inhibition by these inhibitors. Z-VAD-fmk and the peptidomimetic inhibitors inhibit caspase-3 and caspase-8 via a three-step kinetic mechanism. Inhibition of both caspase-3 and caspase-8 by Z-VAD-fmk and of caspase-3 by the peptidomimetic inhibitors proceeds via two rapid equilibrium steps followed by a relatively fast inactivation step. However, caspase-8 inhibition by the peptidomimetics goes through a rapid equilibrium step, a slow-binding reversible step, and an extremely slow inactivation step. The crystal structures of inhibitor complexes of caspases-3 and -8 validate the design of the inhibitors by illustrating in detail how they mimic peptide substrates. One of the caspase-8 structures also shows binding at a secondary, allosteric site, providing a possible route to the development of noncovalent small molecule modulators of caspase activity.},
doi = {10.1016/j.bbapap.2010.05.007},
journal = {BBA- Proteins Proteom.},
issn = {1570-9639},
number = (9) ; 09, 2010,
volume = 1804,
place = {United States},
year = {2010},
month = {9}
}