Biology of Giardia lamblia. Detection of N-acetyl-D-glucosamine as the only surface saccharide moiety and identification of two distinct subsets of trophozoites by lectin binding.
Lectins and glycosidases of known sugar specificity were used as probes to analyze the surface carbohydrate moieties of G. lamblia trophozoites and in particular to determine whether chitin or oligomeric D-GlcNAc is present in the trophozoite form of the parasite as well as on the cyst. Of 13 lectins with varying sugar specificity, only D-GlcNAc-specific lectins bound specifically to the trophozoite surface as determined by light microscopy and EM. A striking finding was the identification of two distinct subsets of trophozoites, distinguished by reactivity with WGA and detected by light microscopy and EM as well as by flow cytometry. Unlike the cyst wall, the trophozoite D-GlcNAc residues were resistant to chitinase treatment. In contrast N-acetyl-beta-D-glucosaminidase abolished WGA binding suggesting that the lectin binds to terminal beta-linked D-GlcNAc residues. These residues were identified as being present on surface glycoproteins by Western blotting of parasite membrane proteins using WGA as a probe. This study identifies D-GlcNAc as the only saccharide moiety detectable by lectin binding on the surface of G. lamblia trophozoites and demonstrates that in contrast to the cyst, chitin is not present in the trophozoite. In addition two distinct subsets of trophozoites were identified based on reactivity with WGA and may represent varying stages of differentiation from trophozoite to cyst.
Ward
H D
HD
Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Tufts University School of Medicine, Boston, Massachusetts.
Alroy
J
J
Lev
B I
BI
Keusch
G T
GT
Pereira
M E
ME
D000117:Acetylglucosamine / Q000032:analysis*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002686:Chitin / Q000032:analysis; D005872:Giardia / Q000032:analysis* / Q000254:growth & development; D005944:Glucosamine / Q000031:analogs & derivatives*; D006023:Glycoproteins / Q000032:analysis; D037102:Lectins / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D006023:Glycoproteins; D037102:Lectins; D008565:Membrane Proteins; D014909:Wheat Germ Agglutinins; D002686:Chitin; D005944:Glucosamine; D000117:Acetylglucosamine