Effect of trypsin and 2-mercaptoethanol on the exposure of sugar residues on the surface of Leishmania donovani chagasi.
Using transmission electron microscopy, gold-labeled lectins, morphometry and enzyme-linked lectin assay, we could show that treatment of promastigotes of Leishmania donovani chagasi with trypsin did not interfere with the binding of lectins (concanavalin A, peanut agglutinin, wheat germ agglutinin and Ricinus communis agglutinin) to the parasite surface. These observations are in agreement with results we previously obtained using a biochemical approach. Treatment of fixed promastigotes with 2-mercaptoethanol induced a significant increase in the density of concanavalin A (Con A) receptors on the surface of L. d. chagasi in relation to the control. We suggest that this increase is due to the unfolding of one or more surface glycoproteins after cleavage of disulfide bonds between cystein residues in adjacent protein loops, exposing second-order Con A receptors that are otherwise hidden in the protein quaternary structure.
Auxiliadora
M
M
Laboratório de Imunopathologia Keizo Asami, Universidade Federal de Pernambuco, Recife, Brazil.
Santos
M
M
de Andrade
P P
PP
de Andrade
C R
CR
Padovan
P A
PA
de Souza
W
W
D000818:Animals; D006023:Glycoproteins / Q000378:metabolism*; D037102:Lectins / Q000378:metabolism; D007893:Leishmania donovani / Q000187:drug effects* / Q000648:ultrastructure; D008623:Mercaptoethanol / Q000494:pharmacology*; D008854:Microscopy, Electron; D011975:Receptors, Mitogen / Q000378:metabolism; D014357:Trypsin / Q000494:pharmacology*
D006023:Glycoproteins; D037102:Lectins; D011975:Receptors, Mitogen; D008623:Mercaptoethanol; D014357:Trypsin