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Mutational analyses of the p35-caspase interaction. A bowstring kinetic model of caspase inhibition by p35.

Title data

Xu, Guozhou ; Rich, Rebecca L. ; Steegborn, Clemens ; Min, Tongpil ; Huang, Yihua ; Myszka, David G. ; Wu, Hao:
Mutational analyses of the p35-caspase interaction. A bowstring kinetic model of caspase inhibition by p35.
In: The Journal of Biological Chemistry. Vol. 278 (14 February 2003) Issue 7 . - pp. 5455-5461.
ISSN 1083-351X
DOI: https://doi.org/10.1074/jbc.M211607200

Abstract in another language

Apoptosis is a highly regulated multistep process for programmed cellular destruction. It is centered on the activation of a group of intracellular cysteine proteases known as caspases. The baculoviral p35 protein effectively blocks apoptosis through its broad spectrum caspase inhibition. It harbors a caspase recognition sequence within a highly protruding reactive site loop (RSL), which gets cleaved by a target caspase before the formation of a tight complex. The crystal structure of the post-cleavage complex between p35 and caspase-8 shows that p35 forms a thioester bond with the active site cysteine of the caspase. The covalent bond is prevented from hydrolysis by the N terminus of p35, which repositions into the active site of the caspase to eliminate solvent accessibility of the catalytic residues. Here, we report mutational analyses of the pre-cleavage and post-cleavage p35/caspase interactions using surface plasmon resonance biosensor measurements, pull-down assays and kinetic inhibition experiments. The experiments identify important structural elements for caspase inhibition by p35, including the strict requirement for a Cys at the N terminus of p35 and the rigidity of the RSL. A bowstring kinetic model for p35 function is derived in which the tension generated in the bowstring system during the pre-cleavage interaction is crucial for the fast post-cleavage conformational changes required for inhibition.

Further data

Item Type: Article in a journal
Refereed: Yes
Additional notes: PubMed-ID: 12458208
Institutions of the University: Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry > Chair Biochemistry - Univ.-Prof. Dr. Clemens Steegborn
Faculties
Faculties > Faculty of Biology, Chemistry and Earth Sciences
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry
Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry
Result of work at the UBT: No
DDC Subjects: 500 Science > 540 Chemistry
Date Deposited: 13 Apr 2015 10:54
Last Modified: 13 Apr 2015 10:54
URI: https://eref.uni-bayreuth.de/id/eprint/10109