Hsp70 mutant proteins modulate additional apoptotic pathways and improve cell survival

Ruiqiong Ran, Guoping Zhou, Aigang Lu, Lu Zhang, Yang Tang, Alan C. Rigby, Frank R Sharp

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Although wild-type Hsp70 (Hsp70WT) inhibits procaspase-3 processing by preventing apoptosome formation, Hsp70WT does not block active caspase-3. Because all caspase-3 inhibitors bear canonical DXXD caspase-3 recognition motifs, we determined whether mutated Hsp70s with caspase-binding motifs act as direct caspase-3 inhibitors. Based on Hsp70 molecular modeling, the DNQP, DEVQ, and EEVD regions localized on the surface of Hsp70WT were chosen, allowing us to design mutants while trying to avoid disrupting the global fold of the molecule and losing the possibility of protein-protein interactions. We replaced DNQP with DQMD, and DEVQ and EEVD with DEVD residues that should be optimal substrates for caspase-3. The resultant Hsp70 mutants directly interacted with active caspase-3 and blocked its proteolytic activity while retaining the ability to reverse protein denaturation and disrupt the interaction between Apaf-1 and procaspase-9. The Hsp70C-terminal mutants interacted with Apaf-1 and active caspase-3 significantly longer than Hsp70WT. The Hsp70 DXXD mutants protected neuron and teratocarcinoma (NT) cells against cell death much better than Hsp70WT whether given before or after serum withdrawal. Hsp70 mutants represent a possible approach to antiapoptotic biotherapeutics. Similar rational designs could be used to engineer inhibitors of additional casoase family members.

Original languageEnglish (US)
Pages (from-to)229-242
Number of pages14
JournalCell Stress and Chaperones
Volume9
Issue number3
DOIs
StatePublished - Sep 2004
Externally publishedYes

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Mutant Proteins
Caspase 3
Cell Survival
Cells
Caspase Inhibitors
Apoptosomes
Teratocarcinoma
Protein Denaturation
Proteins
Denaturation
Aptitude
Molecular modeling
Caspase 9
Cell death
Caspases
Neurons
Cell Death
Engineers
Molecules
Substrates

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Hsp70 mutant proteins modulate additional apoptotic pathways and improve cell survival. / Ran, Ruiqiong; Zhou, Guoping; Lu, Aigang; Zhang, Lu; Tang, Yang; Rigby, Alan C.; Sharp, Frank R.

In: Cell Stress and Chaperones, Vol. 9, No. 3, 09.2004, p. 229-242.

Research output: Contribution to journalArticle

Ran, Ruiqiong ; Zhou, Guoping ; Lu, Aigang ; Zhang, Lu ; Tang, Yang ; Rigby, Alan C. ; Sharp, Frank R. / Hsp70 mutant proteins modulate additional apoptotic pathways and improve cell survival. In: Cell Stress and Chaperones. 2004 ; Vol. 9, No. 3. pp. 229-242.
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