Metacaspase-3 of Plasmodium falciparum: An atypical trypsin-like serine protease

Int J Biol Macromol. 2019 Oct 1;138:309-320. doi: 10.1016/j.ijbiomac.2019.07.067. Epub 2019 Jul 10.


Metacaspases are clan CD cysteine peptidases found in plants, fungi and protozoa that possess a conserved Peptidase_C14 domain, homologous to the human caspases and a catalytic His/Cys dyad. Earlier reports have indicated the role of metacaspases in cell death; however, metacaspases of human malaria parasite remains poorly understood. In this study, we aimed to functionally characterize a novel malarial protease, P. falciparum metacaspase-3 (PfMCA3). Unlike other clan CD peptidases, PfMCA3 has an atypical active site serine (Ser1865) residue in place of canonical cysteine and it phylogenetically forms a distinct branch across the species. To investigate whether this domain retains catalytic activity, we expressed, purified and refolded the Peptidase_C14 domain of PfMCA3 which was found to express in all asexual stages. PfMCA3 exhibited trypsin-like serine protease activity with ser1865 acting as catalytic residue to cleave trypsin oligopeptide substrate. PfMCA3 is inhibited by trypsin-like serine protease inhibitors. Our study found that PfMCA3 enzymatic activity was abrogated when catalytic serine1865 (S1865A) was mutated. Moreover, PfMCA3 was found to be inactive against caspase substrate. Overall, our study characterizes a novel metacaspase of P. falciparum, different from human caspases and not responsible for the caspase-like activity, therefore, could be considered as a potential chemotherapeutic target.

Keywords: Caspase; Chemotherapeutic; Cysteine peptidase; Metacaspase-3; Peptidase_C14; Serine protease.

MeSH terms

  • Amino Acid Sequence
  • Biocatalysis
  • Caspase Inhibitors / pharmacology
  • Caspases / chemistry
  • Caspases / genetics
  • Caspases / metabolism*
  • Catalytic Domain
  • Hydrogen-Ion Concentration
  • Kinetics
  • Plasmodium falciparum / enzymology*
  • Plasmodium falciparum / genetics
  • Serine Endopeptidases / metabolism*
  • Temperature


  • Caspase Inhibitors
  • trypsin-like serine protease
  • Serine Endopeptidases
  • Caspases