High affinity binding of the Entamoeba histolytica lectin to polyvalent N-acetylgalactosaminides.
Entamoeba histolytica trophozoites initiate pathogenic colonization by adherence to host glycoconjugates via an amebic surface lectin which binds to galactose (Gal) and N-acetylgalactosamine (GalNAc) residues. Monovalent and multivalent carbohydrate ligands were screened for inhibition of E. histolytica lectin-mediated human red cell hemagglutination, revealing that: (i) the synthetic multivalent neoglycoprotein GalNAc39BSA (having an average of 39 GalNAc residues linked to bovine serum albumin) was 140,000-fold more potent an inhibitor than monovalent GalNAc and 500,000-fold more potent than monovalent Gal; and (ii) small synthetic multivalent ligands which bind with high affinity to the mammalian hepatic Gal/GalNAc lectin do not bind with high affinity to the E. histolytica lectin. Radioligand binding studies revealed saturable binding of 125I-GalNAc39BSA to E. histolytica membranes (KD = 10 +/- 3 nM, Bmax = 0.9 +/- 0.08 pmol/mg membrane protein). Maximal binding required the presence of calcium chloride (300 microM) or sodium chloride (50 mM), and had a broad pH maximum (pH 6-9). GalNAc39BSA was 200,000-fold more potent than monovalent GalNAc in blocking radio-ligand binding. Among synthetic saccharide-derivatized linear polymers, the GalNAc beta and GalNAc alpha 3Gal beta derivatives were the most potent, with GalNAc alpha and GalNAc alpha 3(Fuc alpha 2)Gal beta derivatives much weaker. The data support a model in which a unique pattern of spaced multiple GalNAc residues are the highest affinity targets for the E. histolytica lectin.
Adler
P
P
Department of Pharmacology and Molecular Science, Johns Hopkins School of Medicine, Baltimore, Maryland 21205.
Wood
S J
SJ
Lee
Y C
YC
Lee
R T
RT
Petri
W A
WA
Schnaar
R L
RL
D000116:Acetylgalactosamine / Q000031:analogs & derivatives* / Q000378:metabolism*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D004748:Entamoeba histolytica / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism; D006001:Glycoconjugates / Q000378:metabolism; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism*
D002241:Carbohydrates; D006001:Glycoconjugates; D037102:Lectins; D008562:Membrane Glycoproteins; C079431:N-acetylgalactosamine-bovine serum albumin conjugate; D015800:Protozoan Proteins; C052452:galactose inhibitable adherence protein, Entamoeba histolytica; D012710:Serum Albumin, Bovine; D000116:Acetylgalactosamine; D005690:Galactose