title: Binding properties of hydrophobic molecules to human serum albumin studied by fluorescence titration.
abstract: Two fluorescence modes were combined to analyze the binding properties of terminally substituted alkanes (C(n)X, X = COOH, OH, CHO, NH(2)) to human serum albumin (HSA). A competitive binding assay using an 8-anilino-1-naphthalenesulfonate (ANS) fluorescence probe provides information on all the hydrophobic binding sites in HSA. A binding assay using the intrinsic fluorescence of the tryptophan residue in HSA (Trp-HSA) provides information on the specific binding site close to the tryptophan residue. There are three fluorescence-active ANS binding sites in HSA, which can be classified into two types by their affinity for ANS. C(n)COOH bound to all three ANS binding sites including the Trp-HSA site, however, it did not quench the fluorescence of Trp-HSA. C(n)CHO bound only to the Trp-HSA site with quenching of the fluorescence of Trp-HSA. By comparing the binding affinities of HSA for C(n)OH and C(n)CHO, it was concluded that the C(n)OH binding site is different from the C(n)CHO binding site. C(n)NH(2) did not bind to any of the three ANS binding sites in HSA.
authors.affiliation: Department of Chemistry, Faculty of Sciences, Kyushu University, Fukuoka 810-8560, Japan. kou@rc.kyushu-u.ac.jp
authors.forename: Kô
authors.initials: K
authors.lastname: Takehara
authors.forename: Keiko
authors.initials: K
authors.lastname: Yuki
authors.forename: Masumi
authors.initials: M
authors.lastname: Shirasawa
authors.forename: Shinya
authors.initials: S
authors.lastname: Yamasaki
authors.forename: Shuto
authors.initials: S
authors.lastname: Yamada
mesh_terms: D000473:Alkanes / Q000737:chemistry*; D000817:Anilino Naphthalenesulfonates; D001665:Binding Sites; D001667:Binding, Competitive; D005456:Fluorescent Dyes; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D015336:Molecular Probe Techniques; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D017186:Titrimetry
chemical_list: C515594:8-anilino-1-naphthalenesulfonic acid; D000473:Alkanes; D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D012709:Serum Albumin