title: Specific cleavage sites on human IgG subclasses by cruzipain, the major cysteine proteinase from Trypanosoma cruzi.
abstract: Cruzipain, the major cysteine proteinase of Trypanosoma cruzi, might have other biological roles than its metabolic functions. In this report, we have explored the interaction of cruzipain with molecules of the immune system. The enzyme was used to digest all human IgG subclasses at different pH values and lengths of time. At pH 7.3, all subclasses were readily split at the hinge region. Immunoblot and amino acid sequence analysis showed fragments of IgG1 and IgG3 to be compatible with Fab and Fc, whereas IgG2 and IgG4 rendered Fab2 and Fc. In all cases the fragments produced might impair the binding capacities and the effector functions of specific IgG. At these cleavage sites cruzipain displays cathepsin L and/or cathepsin B activities and shows a clear preference for Pro at the P'2 position and polar residues at P1. Despite the activity of cruzipain within the hinge, the enzyme also cleaved all heavy chains between the CH2 and CH3 domains; producing Fc'-like-fragments of 14 kDa. These fragments are potential candidates to block or saturate Fc receptors on immunocompetent cells. At mild acidic pH cruzipain produced further degradation of the Fc of all subclasses, the Fd of IgG4 and partially the Fd of IgG1, with the consistent loss of any antibody activity. The L chains apparently were not affected. Thus, cruzipain should be able to modulate, depending on the subclass selected and the pH of the environment, the production and the length of different biologically active/inactive IgG fragments.
authors.affiliation: Unidad de Biología Parasitaria, Facultad de Ciencias, Universidad de la República Oriental del Uruguay, Montevideo, Uruguay.
authors.forename: Patricia
authors.initials: P
authors.lastname: Berasain
authors.forename: Carlos
authors.initials: C
authors.lastname: Carmona
authors.forename: Blas
authors.initials: B
authors.lastname: Frangione
authors.forename: Juan José
authors.initials: JJ
authors.lastname: Cazzulo
authors.forename: Fernando
authors.initials: F
authors.lastname: Goñi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007128:Immunoglobulin Fragments / Q000032:analysis*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins; D014349:Trypanosoma cruzi / Q000201:enzymology*
chemical_list: D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D015800:Protozoan Proteins; D003546:Cysteine Endopeptidases; C063214:cruzipain