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PMID: 10233151 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Trypanosoma cruzi calreticulin is a lectin that binds monoglucosylated oligosaccharides but not protein moieties of glycoproteins.

Molecular biology of the cell ·Vol. 10 ·No. 5 ·1999-05-00 ·Pages 1381-94

Labriola C, Cazzulo JJ, Parodi AJ

Abstract

Trypanosoma cruzi is a protozoan parasite that belongs to an early branch in evolution. Although it lacks several features of the pathway of protein N-glycosylation and oligosaccharide processing present in the endoplasmic reticulum of higher eukaryotes, it displays UDP-Glc:glycoprotein glucosyltransferase and glucosidase II activities. It is herewith reported that this protozoan also expresses a calreticulin-like molecule, the third component of the quality control of glycoprotein folding. No calnexin-encoding gene was detected. Recombinant T. cruzi calreticulin specifically recognized free monoglucosylated high-mannose-type oligosaccharides. Addition of anti-calreticulin serum to extracts obtained from cells pulse-chased with [35S]Met plus [35S]Cys immunoprecipitated two proteins that were identified as calreticulin and the lysosomal proteinase cruzipain (a major soluble glycoprotein). The latter but not the former protein disappeared from immunoprecipitates upon chasing cells. Contrary to what happens in mammalian cells, addition of the glucosidase II inhibitor 1-deoxynojirimycin promoted calreticulin-cruzipain interaction. This result is consistent with the known pathway of protein N-glycosylation and oligosaccharide processing occurring in T. cruzi. A treatment of the calreticulin-cruzipain complexes with endo-beta-N-acetylglucosaminidase H either before or after addition of anti-calreticulin serum completely disrupted calreticulin-cruzipain interaction. In addition, mature monoglucosylated but not unglucosylated cruzipain isolated from lysosomes was found to interact with recombinant calreticulin. It was concluded that the quality control of glycoprotein folding appeared early in evolution, and that T. cruzi calreticulin binds monoglucosylated oligosaccharides but not the protein moiety of cruzipain. Furthermore, evidence is presented indicating that glucosyltransferase glucosylated cruzipain at its last folding stages.

MeSH Terms
1-Deoxynojirimycin/pharmacology Amino Acid Sequence Animals Antibodies/pharmacology Calcium-Binding Proteins/genetics,immunology,metabolism Calreticulin Carbohydrate Sequence Cloning, Molecular Cysteine Endopeptidases/immunology,metabolism Endoplasmic Reticulum/metabolism Enzyme Inhibitors/pharmacology Glycoproteins/chemistry,metabolism Glycoside Hydrolase Inhibitors Glycosylation Hexosaminidases/pharmacology Immune Sera Lectins/drug effects,genetics,metabolism Molecular Sequence Data Oligosaccharides/metabolism Precipitin Tests Protein Folding Protozoan Proteins/drug effects,genetics,metabolism Ribonucleoproteins/genetics,immunology,metabolism Subcellular Fractions Trypanosoma cruzi/chemistry alpha-Glucosidases
Chemicals
Antibodies Calcium-Binding Proteins Calreticulin Enzyme Inhibitors Glycoproteins Glycoside Hydrolase Inhibitors Immune Sera Lectins Oligosaccharides Protozoan Proteins Ribonucleoproteins 1-Deoxynojirimycin 4-nitrophenyl-alpha-glucosidase Hexosaminidases alpha-Glucosidases Cysteine Endopeptidases cruzipain
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Labriola C
Instituto de Investigaciones Bioquímicas Fundación Campomar, 1405 Buenos Aires, Argentina.
Cazzulo J J
Parodi A J
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.10.5.1381
Published
1999-05-00
Pages
1381-94
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25283
Subset
IM
Grants
NIGMS NIH HHS · R01 GM044500 · United States
NIGMS NIH HHS · GM44500 · United States
Databases
GENBANK
AF107115
External Links
PubMed source DOI: 10.1091/mbc.10.5.1381 PMC Full Text View journal details
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