PUBLICATION

Remodeling hydrogen bond interactions results in relaxed specificity of Caspase-3

Authors
Yao, L., Swartz, P., Hamilton, P.T., Clark, A.C.
ID
ZDB-PUB-210116-7
Date
2021
Source
Bioscience Reports   41(1): (Journal)
Registered Authors
Keywords
apoptosis, caspase, enzyme specificity, protein evolution, zebrafish
MeSH Terms
  • Amino Acid Sequence
  • Animals
  • Aspartic Acid/metabolism
  • Caspase 3/chemistry
  • Caspase 3/metabolism*
  • Caspase 6/metabolism
  • Humans
  • Hydrogen Bonding
  • Sequence Homology, Amino Acid
  • Substrate Specificity
  • Valine/metabolism
  • Zebrafish
  • Zebrafish Proteins/chemistry
  • Zebrafish Proteins/metabolism
PubMed
33448281 Full text @ Biosci. Rep.
Abstract
Caspase enzymes play important roles in apoptosis and inflammation, and the non-identical but overlapping specificity profiles (that is, cleavage recognition sequence) direct cells to different fates. Although all caspases prefer aspartate at the P1 position of the substrate, the caspase-6 subfamily shows preference for valine at the P4 position, while caspase-3 shows preference for aspartate. In comparison to human caspases, caspase-3a from zebrafish has relaxed specificity and demonstrates equal selection for either valine or aspartate at the P4 position. In the context of the caspase-3 conformational landscape, we show that changes in hydrogen bonding near the S3 subsite affect selection of the P4 amino acid. Swapping specificity with caspase-6 requires accessing new conformational space, where each landscape results in optimal binding of DxxD (caspase-3) or VxxD (caspase-6) substrate and simultaneously disfavors binding of the other substrate. Within the context of the caspase-3 conformational landscape, substitutions near the active site result in nearly equal activity against DxxD and VxxD by disrupting a hydrogen bonding network in the substrate binding pocket. The converse substitutions in zebrafish caspase-3a result in increased selection for P4 aspartate over valine. Overall, the data show that the shift in specificity that results in a dual function protease, as in zebrafish caspase-3a, requires fewer amino acid substitutions compared to those required to access new conformational space for swapping substrate specificity, such as between caspases-3 and -6.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping