Small-molecule Activators of a Proenzyme

Overview

Journal Science

Specialty Science

Date 2009 Nov 7

PMID 19892984

Citations 75

Authors

Dennis W Wolan

Julie A Zorn

Daniel C Gray

James A Wells

Affiliations

Soon will be listed here.

Abstract

Virtually all of the 560 human proteases are stored as inactive proenyzmes and are strictly regulated. We report the identification and characterization of the first small molecules that directly activate proenzymes, the apoptotic procaspases-3 and -6. It is surprising that these compounds induce autoproteolytic activation by stabilizing a conformation that is both more active and more susceptible to intermolecular proteolysis. These procaspase activators bypass the normal upstream proapoptotic signaling cascades and induce rapid apoptosis in a variety of cell lines. Systematic biochemical and biophysical analyses identified a cluster of mutations in procaspase-3 that resist small-molecule activation both in vitro and in cells. Compounds that induce gain of function are rare, and the activators reported here will enable direct control of the executioner caspases in apoptosis and in cellular differentiation. More generally, these studies presage the discovery of other proenzyme activators to explore fundamental processes of proenzyme activation and their fate-determining roles in biology.

Citing Articles

Prion Protein Endoproteolysis: Cleavage Sites, Mechanisms and Connections to Prion Disease.

Castle A, Westaway D J Neurochem. 2025; 169(1):e16310.

PMID: 39874431 PMC: 11774512. DOI: 10.1111/jnc.16310.

A strategy to disentangle direct and indirect effects on (de)phosphorylation by chemical modulators of the phosphatase PP1 in complex cellular contexts.

Hoermann B, Durr E, Ludwig C, Ercan M, Kohn M Chem Sci. 2024; 15(8):2792-2804.

PMID: 38404380 PMC: 10882499. DOI: 10.1039/d3sc04746f.

Regulation of Peptidase Activity beyond the Active Site in Human Health and Disease.

Obaha A, Novinec M Int J Mol Sci. 2023; 24(23).

PMID: 38069440 PMC: 10707025. DOI: 10.3390/ijms242317120.

The evolution of small molecule enzyme activators.

Dow L, Case A, Paustian M, Pinkerton B, Simeon P, Trippier P RSC Med Chem. 2023; 14(11):2206-2230.

PMID: 37974956 PMC: 10650962. DOI: 10.1039/d3md00399j.

Mucin 1 as a Molecular Target of a Novel Diisoquinoline Derivative Combined with Anti-MUC1 Antibody in AGS Gastric Cancer Cells.

Gornowicz A, Szymanowski W, Bielawski K, Kaluza Z, Michalak O, Bielawska A Molecules. 2021; 26(21).

PMID: 34770912 PMC: 8588261. DOI: 10.3390/molecules26216504.

References

van de Craen M, Declercq W, Van den brande I, Fiers W, Vandenabeele P . The proteolytic procaspase activation network: an in vitro analysis. Cell Death Differ. 1999; 6(11):1117-24. DOI: 10.1038/sj.cdd.4400589. View

Wei M, Zong W, Cheng E, LINDSTEN T, Panoutsakopoulou V, Ross A . Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science. 2001; 292(5517):727-30. PMC: 3049805. DOI: 10.1126/science.1059108. View

Hedstrom L . Serine protease mechanism and specificity. Chem Rev. 2002; 102(12):4501-24. DOI: 10.1021/cr000033x. View

Abdul-Ghani M, Megeney L . Rehabilitation of a contract killer: caspase-3 directs stem cell differentiation. Cell Stem Cell. 2008; 2(6):515-6. DOI: 10.1016/j.stem.2008.05.013. View

Kaufmann S, Earnshaw W . Induction of apoptosis by cancer chemotherapy. Exp Cell Res. 2000; 256(1):42-9. DOI: 10.1006/excr.2000.4838. View

NEURATH H, Walsh K . Role of proteolytic enzymes in biological regulation (a review). Proc Natl Acad Sci U S A. 1976; 73(11):3825-32. PMC: 431226. DOI: 10.1073/pnas.73.11.3825. View

Shi Y . Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell. 2002; 9(3):459-70. DOI: 10.1016/s1097-2765(02)00482-3. View

Blanc C, Deveraux Q, Krajewski S, Janicke R, Porter A, Reed J . Caspase-3 is essential for procaspase-9 processing and cisplatin-induced apoptosis of MCF-7 breast cancer cells. Cancer Res. 2000; 60(16):4386-90. View

Lakhani S, Masud A, Kuida K, Porter Jr G, Booth C, Mehal W . Caspases 3 and 7: key mediators of mitochondrial events of apoptosis. Science. 2006; 311(5762):847-51. PMC: 3738210. DOI: 10.1126/science.1115035. View

10.

Wei Y, Fox T, Chambers S, Sintchak J, Coll J, Golec J . The structures of caspases-1, -3, -7 and -8 reveal the basis for substrate and inhibitor selectivity. Chem Biol. 2000; 7(6):423-32. DOI: 10.1016/s1074-5521(00)00123-x. View

11.

Earnshaw W, Martins L, Kaufmann S . Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem. 2000; 68:383-424. DOI: 10.1146/annurev.biochem.68.1.383. View

12.

Gao J, Sidhu S, Wells J . Two-state selection of conformation-specific antibodies. Proc Natl Acad Sci U S A. 2009; 106(9):3071-6. PMC: 2651286. DOI: 10.1073/pnas.0812952106. View

13.

Bertrand R, Solary E, OConnor P, Kohn K, Pommier Y . Induction of a common pathway of apoptosis by staurosporine. Exp Cell Res. 1994; 211(2):314-21. DOI: 10.1006/excr.1994.1093. View

14.

Kaufmann S . Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: a cautionary note. Cancer Res. 1989; 49(21):5870-8. View

15.

Peterson Q, Goode D, West D, Ramsey K, Lee J, Hergenrother P . PAC-1 activates procaspase-3 in vitro through relief of zinc-mediated inhibition. J Mol Biol. 2009; 388(1):144-58. PMC: 2714579. DOI: 10.1016/j.jmb.2009.03.003. View

16.

Putt K, Chen G, Pearson J, Sandhorst J, Hoagland M, Kwon J . Small-molecule activation of procaspase-3 to caspase-3 as a personalized anticancer strategy. Nat Chem Biol. 2006; 2(10):543-50. DOI: 10.1038/nchembio814. View

17.

Janicke R, Sprengart M, Wati M, Porter A . Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem. 1998; 273(16):9357-60. DOI: 10.1074/jbc.273.16.9357. View

18.

Yi C, Yuan J . The Jekyll and Hyde functions of caspases. Dev Cell. 2009; 16(1):21-34. PMC: 2850564. DOI: 10.1016/j.devcel.2008.12.012. View

19.

Srinivasula S, Fernandes-Alnemri T, Zangrilli J, Robertson N, Armstrong R, Wang L . The Ced-3/interleukin 1beta converting enzyme-like homolog Mch6 and the lamin-cleaving enzyme Mch2alpha are substrates for the apoptotic mediator CPP32. J Biol Chem. 1996; 271(43):27099-106. DOI: 10.1074/jbc.271.43.27099. View

20.

Hardy J, Lam J, Nguyen J, OBrien T, Wells J . Discovery of an allosteric site in the caspases. Proc Natl Acad Sci U S A. 2004; 101(34):12461-6. PMC: 514654. DOI: 10.1073/pnas.0404781101. View