Structure, Folding, and Assembly of (Pro)caspase-3. The Role of the Dimer Interface in Active Site Formation.

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Title: Structure, Folding, and Assembly of (Pro)caspase-3. The Role of the Dimer Interface in Active Site Formation.
Author: Walters, Jad Anthony
Advisors: A. Clay Clark, Committee Chair
Carl Hall, Committee Member
Paul Wollenzien, Committee Member
Bob Rose, Committee Member
Abstract: Effector procaspase-3 plays a vital role in carrying out the final steps of programmed cell death, leading to the destruction of the cell. Because dimerization of (pro)caspase-3 is essential for enzyme stability and activity, it is important to understand the structural details of the interactions at the dimer interface. We show a single site in the dimer interface of (pro)caspase-3 can be used to activate or inhibit the enzyme. The results presented here demonstrate activation of procaspase-3, in the absence of intersubunit linker cleavage, which generates a constitutively active, uninhibitable enzyme that is very efficient in killing both healthy and diseased mammalian cells. Moreover, inhibition may also be achieved utilizing the same site and the structural studies reveal two novel pathways of inhibition. Overall, these studies show how the interactions at the dimer interface in procaspase-3 are essential in formation of a competent active site. In addition to the procaspase-3 interface studies, several crystallographic studies are presented which are aimed at elucidating a structure of procaspase-3. Finally, a comprehensive protocol for carrying out equilibrium folding studies and determining conformational stabilities of macromolecules is provided. All together, this work has lead to exciting and novel discoveries in the field of apoptosis, including new mechanisms to selectively manipulate cell death.
Date: 2009-05-07
Degree: PhD
Discipline: Biochemistry

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