Title
Modulation of procaspase-7 self-activation by PEST amino acid residues of the N-terminal prodomain and intersubunit linker
Date Issued
01 January 2017
Access level
metadata only access
Resource Type
journal article
Author(s)
Universidad de São Paulo
Publisher(s)
Canadian Science Publishing
Abstract
Procaspase-7 zymogen polypeptide is composed of a short prodomain, a large subunit (p20), and a small subunit (p10) connected to an intersubunit linker. Caspase-7 is activated by an initiator caspase-8 and-9, or by autocatalysis after specific cleavage at IQAD198↓S located at the intersubunit linker. Previously, we identified that PEST regions made of amino acid residues Pro (P), Glu (E), Asp (D), Ser (S), Thr (T), Asn (N), and Gln (Q) are conserved flanking amino acid residues in the cleavage sites within a prodomain and intersubunit linker of all caspase family members. Here we tested the impact of alanine substitution of PEST amino acid residues on procaspase-7 proteolytic self-activation directly in Escherichia coli. The p20 and p10 subunit cleavage were significantly delayed in double caspase-7 mutants in the prodomain (N18A/P26A) and intersubunit linker (S199A/P201A), compared with the wild-type caspase-7. The S199A/P201A mutants effectively inhibited the p10 small subunit cleavage. However, the mutations did not change the kinetic parameters (kcat/KM) and optimal tetrapeptide specificity (DEVD) of the purified mutant enzymes. The results suggest a role of PEST-amino acid residues in the molecular mechanism for prodomain and intersubunit cleavage and caspase-7 self-activation.
Start page
634
End page
643
Volume
95
Issue
6
Language
English
OCDE Knowledge area
Bioquímica, Biología molecular
Subjects
Scopus EID
2-s2.0-85035309450
PubMed ID
Source
Biochemistry and Cell Biology
ISSN of the container
08298211
Sponsor(s)
J.E.B., and J.A. conducted the literature review process and wrote the manuscript and created illustrations and figures; J.A., M.G.M., and J.M.O. designed and conducted the experiments and analyzed the data. All of the authors read and approved the final manuscript. We are grateful to Dr. Jennifer L. Harris, Jun Li, Cláudio Joazeiro, Robert Damoiseaux, Christine Tumanut, Julie-Ann Gavigan, and Reynard Pacoma of the Genomics Institute of the Novartis Research Foundation, San Diego, California, USA, for outstanding technical support and reagents. This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP 01/01000-7, 08/50051-2) and The Genomics Institute of the Novartis Research Foundation. The authors declare that they have no conflict of interest associated with this study.
Sources of information:
Directorio de Producción Científica
Scopus