Since 1980's, reversed micelles, as a mimetic membrane system, 1 have attracted the increasing interest of chemists and biologists. They can provide a nano-sized reaction room, 2 sometimes entitled reverse micellar microreactors. 3 Reversed micelles have been widely studied concerning enzyme activity, [4][5][6] the solubilization mechanism of protein, 7 conformational changes of protein, 8 the synthesis of nanoparticles, 9-12 and so on. Nanoparticle probes, compared with organic dyes acting as biosensors in chemical and biochemical fields, have been researched recently and their application is becoming more extensive. Three types of nanoparticles in biochemical analysis are used: metal nanoparticles, 13 silica nanoparticles, 14,15 and luminescence quantum dot. 16,17 These probes have been applied to the ultrasensitive detection of proteins, DNA sequencing, clinical diagnostics, etc. In the case of in vivo studies, the nanoparticles will possibly affect the tissues or cells of an organism. For example, Cd 2+ and S 2-are component parts of a quantum dot. Also, as we know, the two ions are harmful to the human body. By researching the interaction of CdS nanoparticles with BSA in reversed micelles, the effect of CdS to protein in real cells can be simulated.In this work, the effects of the microenvironment induced by reversed micelles on the fluorescence spectra of BSA were investigated. The focus of this work was researching the effects of neutral and charged CdS nanoparticles on BSA-fluorescence quenching in reversed micelles. The quenching may be the result of energy transfer between BSA and CdS nanoparticles. A quenching equation according to the static quenching model and the quenching constant was obtained.
Experimental
ApparatusFluorescence and UV spectra were measured on a Hitachi F-4500 fluorescence spectrophotometer and a TU-1221 PUXI General UV-VIS spectrophotometer, respectively.
ReagentsAll chemicals were analytical-grade regents. Doubly distilled water via a double-distillation quartz set was used throughout.Bovine serum albumin (BSA), purchased from LiZhu DongFeng Biotechnology, Co. LTD. (Shanghai, China), was used without further purification. A stock solution of BSA at a concentration of 30 mg ml -1 was prepared using water and stored at 4˚C prior to use. A cetyltrimethylammonium bromide (CTAB) solution (0.05 mol L -1 ) was prepared by dissolving an appropriate amount of CTAB into 500 ml of 1-butanol-cyclohexane mixture (v/v = 1/9), where 1-butanol acted as a co-surfactant.
ProcedureAppropriate quantities of BSA aqueous solution and water were transferred to a small flask using a microsyringe, followed by adding a CTAB organic solution to a total volume of 10.00 ml. The water content, ω (= [H2O]/[CTAB]), was adjusted by changing the volume of water at a fixed BSA concentration in reversed micelles. A blank system was prepared similarly, but by replacing the BSA solution with water of the same volume. Both the BSA-H2O-CTAB mixtures and the blank were ultrasonicated for 5 min until the micellar suspen...