湖北大学学报（自然科学版）
湖北大學學報（自然科學版）
호북대학학보（자연과학판）
2015年
3期
282-286,291
,共6页
7-(8-喹啉)偶氮-8-羟基喹啉-5-磺酸%BSA%HSA%荧光光谱
7-(8-喹啉)偶氮-8-羥基喹啉-5-磺痠%BSA%HSA%熒光光譜
7-(8-규람)우담-8-간기규람-5-광산%BSA%HSA%형광광보
8-hydroxy-7-(quinolin-8-ylazo)-quinoline-5-sulfonic acid%BSA%HSA%fluorescence spectrometry
应用紫外吸收光谱法和荧光光谱法研究了7-[（8-喹啉）偶氮]-8-羟基喹啉-5-磺酸（简称主体）与蛋白质的相互作用.在pH=7.40的Tris-HCl缓冲溶液中，主体分子在400 nm处发射弱荧光，其荧光强度随白蛋白的加入明显增强.据此建立测定微量蛋白质的新方法.在相同的实验条件下，测定牛血清白蛋白和人血清白蛋白，线性范围分别为1.0×10-6~2.7×10-5 mol/L和2.5×10-7~9.5×10-6 mol/L，检测限分别为8.09×10-7 mol/L、1.05×10-7 mol/L.同时讨论7-[（8-喹啉）偶氮]-8-羟基喹啉-5-磺酸对白蛋白内源荧光的猝灭机理，测定牛血清白蛋白和人血清白蛋白结合常数分别为3.38×105和7.28×106，结合位点数分别为1.218和1.412.依据Forster非辐射能量转移理论，确定了主体-受体间的结合距离rBSA=3.98 nm，rHSA=3.04 nm及能量转移效率EBSA=0.038，EHSA=0.161，用同步荧光技术考察主体对蛋白质构象的影响.①
應用紫外吸收光譜法和熒光光譜法研究瞭7-[（8-喹啉）偶氮]-8-羥基喹啉-5-磺痠（簡稱主體）與蛋白質的相互作用.在pH=7.40的Tris-HCl緩遲溶液中，主體分子在400 nm處髮射弱熒光，其熒光彊度隨白蛋白的加入明顯增彊.據此建立測定微量蛋白質的新方法.在相同的實驗條件下，測定牛血清白蛋白和人血清白蛋白，線性範圍分彆為1.0×10-6~2.7×10-5 mol/L和2.5×10-7~9.5×10-6 mol/L，檢測限分彆為8.09×10-7 mol/L、1.05×10-7 mol/L.同時討論7-[（8-喹啉）偶氮]-8-羥基喹啉-5-磺痠對白蛋白內源熒光的猝滅機理，測定牛血清白蛋白和人血清白蛋白結閤常數分彆為3.38×105和7.28×106，結閤位點數分彆為1.218和1.412.依據Forster非輻射能量轉移理論，確定瞭主體-受體間的結閤距離rBSA=3.98 nm，rHSA=3.04 nm及能量轉移效率EBSA=0.038，EHSA=0.161，用同步熒光技術攷察主體對蛋白質構象的影響.①
응용자외흡수광보법화형광광보법연구료7-[（8-규람）우담]-8-간기규람-5-광산（간칭주체）여단백질적상호작용.재pH=7.40적Tris-HCl완충용액중，주체분자재400 nm처발사약형광，기형광강도수백단백적가입명현증강.거차건립측정미량단백질적신방법.재상동적실험조건하，측정우혈청백단백화인혈청백단백，선성범위분별위1.0×10-6~2.7×10-5 mol/L화2.5×10-7~9.5×10-6 mol/L，검측한분별위8.09×10-7 mol/L、1.05×10-7 mol/L.동시토론7-[（8-규람）우담]-8-간기규람-5-광산대백단백내원형광적졸멸궤리，측정우혈청백단백화인혈청백단백결합상수분별위3.38×105화7.28×106，결합위점수분별위1.218화1.412.의거Forster비복사능량전이이론，학정료주체-수체간적결합거리rBSA=3.98 nm，rHSA=3.04 nm급능량전이효솔EBSA=0.038，EHSA=0.161，용동보형광기술고찰주체대단백질구상적영향.①
The binding characteristics of 8-hydroxy-7-(quinolin-8-ylazo)-quinoline-5-sulfonic acid (host)and serum albumin were studied by fluorescence and absorption spectroscopy in aqueous solution.The host gave fluorescence emission at 400 nm(λex=320 nm)in a Tris-HCl buffer solution (pH=7.40). A new method for the determination of trace protein HSA has been developed based on the enhanced fluorescence emission in the presence of a certain amount of protein. Under the same conditions the liner range concentration was at 1.0×10-6 2.7×10-5 mol/L and 2.5×10-7 9.5×10-6 mol/L,for bovine serum albumin and human serum albumin,respectively. The detection limit was 8.09×10-7 mol/L,1.05×10-7 mol/L.At the same time,the results show that the binding constants,the number of binding sites and the quenching mechanism of fluorescence of serum albumin by host indicate a static quenching procedure for bovine serum albumin and human serum albumin.The binding constants is 3.38 × 105 and 7.28 × 106,the number of binding sites is 1.218 and 1.412. The binding distance between host(rBSA=3.98 nm,rHSA=3.04 nm) and serum albumin and the energy transfer efficiency(EBSA=0.038,EHSA=0.161)are obtained based on the theory of Forester spectroscopy energy transfer. The effect of host on the conformation of serum albumin is further analyzed by synchronous fluorescence spectrometry.
