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Trypanosoma cruzi: cruzipain and membrane-bound cysteine proteinase isoform(s) interacts with human

Adrián M Ramos1, Vilma G Duschak, Nelia M Gerez de Burgos

  • 1Departamento de Bioquímica Clínica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, (5000), Argentina. amramos@bioclin.fcq.unc.edu.ar

Experimental Parasitology
|June 11, 2002
PubMed

Insights

Human alpha-macroglobulin (alpha-M) inhibitors, pregnancy zone protein (PZP) and alpha2-macroglobulin (alpha2-M), bind to Trypanosoma cruzi proteinases. This interaction inhibits cruzipain activity, potentially protecting the host from parasitic damage.

Area of Science:

  • Biochemistry
  • Parasitology
  • Immunology

Background:

  • Plasmatic levels of pregnancy zone protein (PZP) increase in children with acute Chagas disease.
  • PZP and alpha2-macroglobulin (alpha2-M) are known to interact with Trypanosoma cruzi proteinases.

Purpose of the Study:

  • Investigate the interaction between alpha2-M, PZP, and cruzipain, the major cysteine proteinase of T. cruzi.
  • Determine the molecular changes in alpha-M inhibitors upon reaction with cruzipain.
  • Assess the impact of this interaction on cruzipain activity and potential host protection.

Main Methods:

  • Polyacrylamide gel electrophoresis (PAGE) analysis to detect complex formation, proteolysis, and covalent binding.
  • Gelatin SDS-PAGE and immunoblotting using anti-cruzipain antibodies to detect cruzipain-alpha-M complexes.
  • Enzyme activity assays using Bz-Pro-Phe-Arg-pNA substrate to measure cruzipain inhibition.

Main Results:

  • PAGE analysis revealed complex formation, tetramerization of PZP, limited proteolysis of alpha-M inhibitors, and covalent binding of cruzipain to PZP and alpha2-M.
  • Conformational and structural changes in alpha-M inhibitors correlated with modified enzyme mobility and activity.
  • Cruzipain-alpha-M complexes were confirmed, and alpha2-M and PZP significantly impaired cruzipain activity.
  • Alpha-M inhibitors also formed covalent complexes with membrane-bound cysteine proteinases cross-reacting with cruzipain.

Conclusions:

  • Human alpha-macroglobulin inhibitors, alpha2-M and PZP, interact with and inhibit the activity of T. cruzi cruzipain.
  • These inhibitors may protect host molecules from cruzipain-mediated damage.
  • The interaction could potentially regulate parasite functions controlled by cruzipain, offering a novel therapeutic avenue.

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