Identification and characterization of taglin, a mannose 6-phosphate binding, trypsin-activated lectin from Giardia lamblia.
We have previously reported the presence of a cell surface associated lectin activity in Giardia lamblia, a human protozoan parasite that is a significant cause of diarrheal disease worldwide [Lev, B., Ward, H., Keusch, G. T., & Pereira, M. E. A. (1986) Science (Washington, D.C.) 232, 71-73]. This lectin is specifically activated in vitro by a host protease, trypsin, which is secreted in vivo at the site of infection. The activated lectin agglutinates cells to which the parasite adheres in vivo and binds specifically to isolated brush border membranes of these cells. These findings suggest that this lectin may be of importance in the host-parasite interaction. We now report the identification of this lectin, which we have named taglin (to denote trypsin-activated Giardia lectin), and describe some of its properties. A monoclonal antibody that inhibits the hemagglutinating activity of taglin recognizes a protein of 28,000/30,000 kdaltons in Western blots of Giardia lysates. This finding was confirmed by direct demonstration of lectin activity with the technique of erythrocyte binding to proteins electroblotted to nitrocellulose, which revealed specific red cell binding to giardial protein bands in the same molecular weight range as those recognized by the monoclonal antibody. This study also elucidates the binding of taglin to terminal phosphomannosyl residues. The involvement of cell surface phosphate in binding of taglin to erythrocytes is shown by the abolition of lectin activity by alkaline phosphatase treatment of the erythrocytes. Taglin also requires divalent cations, Ca2+ or Mn2+, for hemagglutinating activity and is active within a narrow pH range of 6-7.
Ward
H D
HD
Division of Geographic Medicine and Infectious Diseases, New England Medical Center Hospitals, Tufts University School of Medicine, Boston, Massachusetts 02111.
Lev
B I
BI
Kane
A V
AV
Keusch
G T
GT
Pereira
M E
ME
D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D002241:Carbohydrates; D002413:Cations, Divalent; D004912:Erythrocytes / Q000276:immunology; D005872:Giardia / Q000276:immunology*; D006384:Hemagglutination; D037102:Lectins / Q000302:isolation & purification*; D008360:Mannosephosphates; D015800:Protozoan Proteins; D014357:Trypsin / Q000494:pharmacology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D002241:Carbohydrates; D002413:Cations, Divalent; D037102:Lectins; D008360:Mannosephosphates; D015800:Protozoan Proteins; C055413:taglin protein, Giardia lamblia; C027693:mannose-6-phosphate; D014357:Trypsin