title: Characterization of the adenine binding sites of two Dolichos biflorus lectins.
abstract: The seed lectin and a stem and leaf lectin (DB58) from Dolichos biflorus have high-affinity hydrophobic sites that bind to adenine. The present study employs a centrifugal filtration assay to characterize these sites. The seed lectin contains two identical sites with Ka's of 7.31 x 10(5) L/mol whereas DB58 has a single site with a Ka of 1.07 x 10(6) L/mol. The relative affinities of these sites for a host of adenine analogs and derivatives were determined by competitive displacement assays. The most effective competitors for adenine were the cytokinins, a class of plant hormone, for which the lectins had apparent Ka's of 1.96 x 10(5)-4.90 x 10(4) L/mol. Direct binding of the cytokinin 6-(benzylamino)purine (BAP) to both lectins showed positive cooperativity for only the seed lectin, indicating the interaction of this ligand with more than one class of hydrophobic binding site. Fluorescence enhancement assays demonstrate cooperativity between hydrophobic sites of the seed lectin and also suggest that BAP binds to more than one class of site.
authors.affiliation: Department of Biochemistry and Biophysics, University of California, Davis 95616.
authors.forename: C V
authors.initials: CV
authors.lastname: Gegg
authors.forename: D D
authors.initials: DD
authors.lastname: Roberts
authors.forename: I H
authors.initials: IH
authors.lastname: Segel
authors.forename: M E
authors.initials: ME
authors.lastname: Etzler
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism*; D000817:Anilino Naphthalenesulfonates; D001593:Benzyl Compounds; D001665:Binding Sites; D001667:Binding, Competitive; D003583:Cytokinins / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes; D007701:Kinetin; D037102:Lectins / Q000378:metabolism*; D037121:Plant Lectins; D011687:Purines; D014462:Ultrafiltration
chemical_list: D000817:Anilino Naphthalenesulfonates; D001593:Benzyl Compounds; D003583:Cytokinins; D005456:Fluorescent Dyes; D037102:Lectins; D037121:Plant Lectins; D011687:Purines; C036554:dolichos biflorus agglutinin; C027132:1-anilino-8-naphthalenesulfonate; D000225:Adenine; C480551:benzylaminopurine; D007701:Kinetin