title: Crosslinking of low-affinity glycoprotein ligands to galectin LEC-1 using a photoactivatable sulfhydryl reagent.
abstract: The N-terminal lectin domain (Nh) of the tandem repeat-type nematode galectin LEC-1 has a lower affinity for sugars than the C-terminal lectin domain. To confirm that LEC-1 forms a complex with N-acetyllactosamine-containing glycoproteins, we used several mutants of LEC-1 in which a unique cysteine residue was introduced into the Nh domain and examined their binding to bovine asialofetuin with a photoactivatable sulfhydryl crosslinking reagent. A crosslinked product was formed with the Q38C mutant, strongly suggesting the low-affinity interaction of Nh with the glycoprotein could be detected with this system.
authors.affiliation: Department of Biological Chemistry, Teikyo University School of Pharmaceutical Sciences, Sagamiko, Kanagawa 199-0195, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Arata
authors.forename: Mayumi
authors.initials: M
authors.lastname: Tamura
authors.forename: Takamasa
authors.initials: T
authors.lastname: Nonaka
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Kasai
mesh_terms: D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D003432:Cross-Linking Reagents / Q000737:chemistry*; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D060748:Fetuins; D037483:Galectin 1 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D008024:Ligands; D009154:Mutation / Q000235:genetics; D010777:Photochemistry; D013439:Sulfhydryl Reagents / Q000737:chemistry*; D000509:alpha-Fetoproteins / Q000737:chemistry / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001212:Asialoglycoproteins; D003432:Cross-Linking Reagents; D060748:Fetuins; D037483:Galectin 1; D006023:Glycoproteins; D008024:Ligands; D013439:Sulfhydryl Reagents; D000509:alpha-Fetoproteins; C017029:asialofetuin; D001616:beta-Galactosidase; D003545:Cysteine