Binding Properties of Hydrophobic Molecules to Human Serum Albumin Studied by Fluorescence Titration

Ko Takehara, Keiko Yuki, Masumi Shirasawa, Shinya Yamasaki, Shuto Yamada

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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.

Original languageEnglish
Pages (from-to)115-120
Number of pages6
Journalanalytical sciences
Issue number1
Publication statusPublished - Jan 1 2009


All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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