Classification of the binding modes in bovine serum albumin using terminally substituted alkane analogues.
With the fluorescence probe of 8-anilino-1-naphthalenesulfonate (ANS), the binding modes of terminally substituted alkane analogues (C(n)X; X = COOH, OH, CHO, NH(3), CONH(2)) to bovine serum albumin (BSA) were investigated using a competitive binding technique. The Scatchard plot of the fluorometric titration of BSA with ANS showed that the maximum binding number of ANS, n(max), was 3.81, with the binding constant, K(bnd), of 1.42 x 10(6) mol(-1) dm(3). The binding modes of C(n)X to BSA were analyzed based on the fluorometric titration of the ANS and BSA mixture with C(n)X. C(n)COOH completely displaced the ANS bound to BSA, whereas C(n)OH and C(n)CHO displaced only about 40% of the ANS bound to BSA. In contrast, C(n)NH(2) and C(n)CONH(2) displaced very little bound ANS. By comparing these results, we classified the binding modes of C(n)X to BSA into three types. Two of them are detectable with the ANS fluorescence and the remaining one is not detectable with the fluorescence.
Takehara
Kô
K
Department of Chemistry, Faculty of Sciences, Kyushu University, Fukuoka, Japan. kou@rc.kyushu-u.ac.jp
Morinaga
Yuki
Y
Nakashima
Shinya
S
Matsuoka
Shiro
S
Kamaya
Hiroshi
H
Ueda
Issaku
I
D000473:Alkanes / Q000737:chemistry*; D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D002965:Classification; D005456:Fluorescent Dyes; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D017186:Titrimetry
D000473:Alkanes; D005456:Fluorescent Dyes; D012710:Serum Albumin, Bovine