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1 Vol. 27 No CHEM ICAL JOURNAL OF CH INESE UN IVERSITIES ,, (,, ),, ( 2 ) ;, HSA BSA, 2,. ; (HSA) ; (BSA) ; O644; O657 A (2006) ( ) ( ) ( ) ( ),. DNA, DNA [ ], [ 2, 3 ], DNA [ 4 ].,.,,, [ 5 3 ],,.., [ 5, 0, ]. [ 6 ] [ 7 ], [ 3 ],,,.. (HSA ) (BSA ), ; ( ) ; ( ) ; ( ) ( )..,.. 2 HSA BSA 005 mol/l Tris2HCl (ph = 74), mol/l. RF2540, Tris2HCl, cm, 280 nm, nm, : : ( : ) ( : ). : (933 ),,,,. E2mail: yangp in@ sxu. edu. cn

2 688 Vol. 27. UV2265 n ( ) n ( ) = ( ) nm n [ 6 ] : F 0 F = + K A c (Q ) () F 0 - F = K D + F 0 F 0 c (Q ), c (Q ), F 0 F. ( 2) ( F 0 - F) - c (Q ) -, K D. 2. 2,. [ 7 ] : ( F - F A ) M t M b = = K A ( np t - M b ) ( F b - F A ) + (2) + (3) K A ( np t - M b ), M t, M b, P t, M n P M b / n, M =M t - M b, P = P t - (M b / n). F = F - F D [ F, F D ( ), F A ( ), F b ]. (4), ( np t ( F - F A ) -,, K A n (4) - M b ) - F rster 2, E 2 r R 0 J, [ 9 ] ( 22-8) - (22-2). 3 3., HSA 330 nm., HSA. HSA, F 0 / F, HAS c (Q ) / c ( P t ),, HSA. BSA, HSA, BSA 330 nm., BSA, BSA 2. HSA,, HSA,,. BSA ( 2) : BSA F ig. The quench ing curve of L evofloxac in Hydrochlor ide to HSA

3 No. 4 : 689 F ig. 2 The quench ing curves of four k inds of m ed ic ine to BSA (A) Levofloxacin hydrochloride; (B) Worfloxacin; (C) Fleroxacin; (D) Gatifoxacin., BSA 4, 2 [ 3 ], ; BSA,, 3, 4 5,, 2 [ 3 ]. (2) HSA BSA,, HSA BSA 3 (A ) (B ). F ig. 3 The double rec iproca l curve of the action of L evofloxac in Hydrochlor ide w ith HSA( A) and L evofloxac in Hydrochlor ide w ith BSA( B) ba sed on fluorescence quench ing form ula( 2) 3 ( ). M t / P t ( ) ( ),,. J, R 0, E, 2 r M t / P t. Table D issoc ia tion con stan t of the four k inds of m ed ic ines w ith HSA( upper) and BSA( nether) K D, linear in2 terrela tion coeff ic ien t C, quench ing con stan t K q, energy tran sfer eff ic iency E, action d istance R 0 and r ( ba sed on fluorescence quench ing form ula( 2) and form ula of d istance of donor2acceptor from [ 9] ) No. D rug K D / (mol L - ) K q / (mol - L - s - ) C 0-4 J R 0 /nm E r/nm M t /P t Levofloxacin Worfloxacin Fleroxacin Galifoxacin ( ) [ 6 ] Stern2Volmer [ 8 ] F 0 / F = + K q 0 c (Q ), K A = / K D = K q 0,, 0 0 ns [ 8 ], K D HSA BSA K q. [ 8 ] [ (mol/l) - s - ]., K q,.

4 690 Vol HSA BSA,, (4),, HSA BSA 4 (A ) (B ). F ig. 4 The double rec iproca l curve of the action of L evofloxac in Hydrochlor ide and HSA( A) L evofloxac in Hydrochlor ide and BSA( B) ba sed on fluorescence enhancem en t form ula( 4) (4), 4 (A) (B) HSA BSA 2. Table 2 D issoc ia tion con stan t of the four k inds of m ed ic ine w ith HSA( upper) and BSA( nether) K D, linear in terre2 la tion coeff ic ien t C, quench ing con stan t K q, energy tran sfer eff ic iency E, action d istance R 0 and r 3 No. D rug K D / (mol L - ) K q / (mol - L - s - ) C 0-4 J R 0 /nm E r/nm M t /P t Levofloxacin Worfloxacin Fleroxacin Gatifoxacin Based on fluorescence enhancement formula (4) and formula of distance of donor2accep tor from [ 9 ] K D,.,,. (2) (4). K D,,,,. BSA ( 2), BSA, BSA, BSA, 4 ; BSA,, 3 5,, 2 [ 2 ]., 2 BSA,. HSA, BSA,, HSA. M t / P t,, 8 ( BSA) 3 ( HSA ),., [ 9 ]

5 No. 4 : 69,,, F rster 2. HSA BSA,, HSA BSA 30% 29%, 20%.. [ ] Skauge T., Turel I., Sletten Einar. Inorganica Chim ica Acta[ J ], 2002, 339: [ 2 ] Kamp ranis S. C., Maxwell A.. J. B iol. Chem. [ J ], 998, 273: [ 3 ] Igor D. V ilfan, Petra D reven Sek, Iztok Turel et al.. B iochim ica et B iophysica Acta[ J ], 2003, 628: 22 [ 4 ] Son G. S., Yeo J. A., Kim M. S. et al.. J. Am. Chem. Soc. [ J ], 998, 20: [ 5 ] ZHU Keng( ), TONG Shen2Yang ( ). Chem. J. Chinese Universities ( ) [ J ], 996, 7 ( 4 ) : [ 6 ] YANG Man2Man ( ), YANG Pin ( ), ZHANG L i2w ei ( ). Chinese Science Bulletin ( ) [ J ], 994, 39 (9) : [ 7 ] Yang P., YangM. M., Yang B. S.. Chinese Journal of Chem istry[ J ], 996, 4 (2) : 09 3 [ 8 ] YANG Pin ( ). Introduction of B ioinorganic Chem istry( ) [M ], Xian: Xian Jiaotong University Press, 99 [ 9 ] YANG Pin ( ), GAO Fei( ). Princip le of B ioinorganic Chem istry ( ) [M ], Beijing: Science Press, 2002 [ 0 ] ZHU Keng( ), TONG Shen2Yang( ). Acat Chim ica Sinica ( ) [ J ], 997, 55 (4) : [ ] ZHU Keng( ), TONG Shen2Yang( ). Chinese Journal of Analytical Chem istry ( ) [ J ], 996, 24 ( ) : [ 2 ] YANG B in2sheng( ), YANG Pin ( ). Progress in B iochem istry and B iophysics( ) [ J ], 992, 9 (2) : 0 4 [ 3 ] YANG Man2Man ( ), X I Xiao2L i( ), YANG Pin ( ). Chinese Science Bulletin ( ) [ J ], 2005, 50 ( 22) : Stud ies on In teraction of New M ed ic ine of Qu inolone2c la ss w ith HSA and BSA by Using Fluorescence Enhancem en t and Fluorescence Quench ing Theory YANG Man2Man, X I Xiao2L i, YANG Pin 3 ( Key L aboratory of Chem ical B iology and M olecular Engineering of M inistry of Education, Institute of M olecular Science, College of Chem istry and Chem ical Engineering, Shanxi U niversity, Taiyuan , China) Abstract By using the two theories fluorescence enhancement and fluorescence quenching, we studied and compared the inter action of four kinds of Quinolone2class2medicines w ith HSA and BSA, then analysed deep ly the binding speciality of m edicine and album in and usual characterization constants: dissociation con2 stant, quenching constant, quenching efficiency, energy2transfer efficiency, the distance between donor and accep ter. About binding type of album in w ith medicine, through the quenching experiment result of the four kinds of medicines with HSA and BSA, we found that this quenching action type caused by the binding of do2 nor between accep tor was not determ ined by the oneself2biomacromolecule B SA, but together determ ined by the molecular structure of BSA and m edicine, donor and accep tor, and their match w ith each other. Keywords Fluorescence enhancement and quenching; HSA; BSA; Quinolone2class2medicine ( Ed. : D, I)