Interaction between bovine serum
albumin and Indo-1 using fluorescence spectroscopic method

doi:10.1007/s11458-008-0013-4

Front. Chem. China

2008, Vol. 3

Issue (1) : 105-111 https://doi.org/10.1007/s11458-008-0013-4

Interaction between bovine serum albumin and Indo-1 using fluorescence spectroscopic method

Haixin BAI¹, BAI Haixin², YANG Cheng², YANG Xiurong²

1.Graduate University of Chinese Academy of Sciences; 2.State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences

Download: PDF(152 KB) HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

Abstract This work attempts to calculate the binding-site number using fluorescence spectroscopic method with bovine serum albumin (BSA) and Indo-1 as protein and ligand models, respectively. The method for calculating the binding-site number in BSA for Indo-1 was developed based on the relationships between changes in Indo-1 fluorescence intensity and the analytical concentration of BSA. The interaction between BSA with Indo-1 was investigated comprehensively using fluorescence techniques as well as fluorescence resonance energy transfer, and the thermodynamic parameters were calculated according to the effect of enthalpy on temperature. Three binding sites in BSA for Indo-1 were revealed, and the distances from Trp212 in BSA to the three binding sites were 2.93, 2.57 and 2.40 nm, respectively. It was also proven that Indo-1 embedded into the three hydrophobic cavities of BSA by hydrophobic association. This paper provides a reference on calculating the binding-site number in proteins for ligands and studying their interactions by fluorescence spectroscopic methods. In fluorescent quenching experiments, fluorescence changes were automatically recorded in real time by combining the Microlab 500 Series Dispenser and PTI fluorescence apparatus.

Issue Date: 05 March 2008

Cite this article:

BAI Haixin,Haixin BAI,YANG Cheng, et al. Interaction between bovine serum albumin and Indo-1 using fluorescence spectroscopic method[J]. Front. Chem. China, 2008, 3(1): 105-111.

URL:

https://academic.hep.com.cn/fcc/EN/10.1007/s11458-008-0013-4
https://academic.hep.com.cn/fcc/EN/Y2008/V3/I1/105

1	Lakowicz J R Principlesof Fluorescence Spectroscopy (2nd Edition)Kuwer Acdemic/Plenum PublisherNew York 1999 ap1bp239cp248dp240
2	Romer J Bichkel M H Method to estimate bindingconstants at variable protein concentrationsJ Pharm Pharmacol 1979 31711
3	Bhattacharyya M Chaudhuri U Poddar R K Evidence for Cooperative Binding of Chloropromazine WithHemoglobin: Equilibrium Dialysis, Fluorescence Quenching and OxygenRelease StudyBiochem Biophys Res Commun 1990 16711461153
4	Zhao H Su W Luo Y H Ji Y H Li ZC. Jiu H F Liang H Chen B Zhang Q J Rectificationof excitation with bathochromic shift induced by intense absorptionof organic ligands during emission. SpectrochimActa A 2006 65846851
5	Subbiah D Ashok K M Fluorescence spectroscopicstudy of serum albumin-bromadiolone interaction: fluorimetric determinationof bromadioloneJ Pharm Biomed Anal 2005 38556563
6	Hirshfield K M Toptygin D Grandhige G Kim H Packard B Z Brand L Steady-stateand time-resolved fluorescence measurements for studying molecularinteractions: interaction of a calcium-binding probe with proteinsBiophys Chem 1996 622538
7	Richieri G V Anel A Kleinfeld A M Interaction of long-chain fatty acids and albumin: determinationof free fatty acid levels using the fluorescent probe ADIFABBiochemistry 1993 3275747580
8	Olson M K Hollingworth S Baylor S M Myoplasmic binding of fura-2 investigated by steady-statefluorescence and absorbance measurementsBiophys J 1988 5410891104
9	Kurebayashi N Harkins A B Baylor S M Use of fura red as an intracellular calcium indicator infrog skeletal muscle fibersBiophys J 1993 6419341960
10	Cantor C R Schimmel P R Biophysical Chemistry, PartIII: The Behavior of Biological Macromolecules (Freeman, San Francisco) 1980 pp. 849886
11	Yang. Yang. Zhang. Studies on the interaction of caffeinic drugs with albuminby fluorescence methodsChinese Sci Bull 1994 39(1)3135
12	Vallner J J Bindingof drugs by albumin and plasma-proteinJPharm Sci 1997 66447465
13	Carter D C Ho J X Structure of serum albuminAdv Protein Chem 1994 45153203
14	Ikenouchi H Peeters G A Barry W H Evidence that binding of indo-1 to cardiac myocyte proteindoes not markedly change Kd for Ca²⁺ CellCalciumCell Calcium 1991 12415422
15	Owen C S Shuler R L Spectral evidence for non-calciuminteractions of intracellular indo-1BiochemBiophys Res Commun 1989 163328333
16	Bancel F Salmon J M Vigo J Viallet P Microspectrofluorometryas a tool for investigation of noncalcium interactions of indo-1Cell Calcium 1992 135968
17	Grynkiewicz G Poenie M Tsien R Y A new generation of Ca²⁺ indicatorswith greatly improved fluorescence propertiesJ Biol Chem 1985 260(6)34403450
18	Ribou A C Vigo J Viallet P Interaction of a protein, BSA, and a fluorescent probe,Mag-Indo-1, influence of EDTA and calcium on the equilibriumJ M Salmon, Biophys Chem 1999 81179189
19	Morelle B Salmon J M Vigo J Viallet P Measurement ofintracellular magnesium concentrations in 3T3 fibroblasts with the.fluorescent indicator MagIndo-1Anal Biochem 1994 218170176
20	Tian J Liu J Tian X Hu Z Chen X Study of the interaction of kaernpferolwith bovine serum albuminJ Mol Struct 2004 691197202
21	Philip D Ross Sabramarian S Thermodynamics of protein associationreactions: forces contributing to stabilityBiochemistry 20 (1981) 30963102
22	Krang-hansen U Molecularaspects of ligand binding to serum albuminPharmacol Rev 1981 33(1)1753
23	Wei Y J Li K A Tong S Y The interaction of Bromophenol blue with proteins in acidicsolutionTalanta 1996 43110
24	Yang B Spectral studies on the combination of lanthanonion with partner albumin. Chem J Chinese U 1998 19(7)10571061
25	Chi Y Zhuang J Li N Li K Tong S Studies on the interaction mechanism ofbovine serum albumin with zinc reagentChemJ Chinese U 1999 20(11)16971702
26	Förster T Modern Quantum Chemistry, vol. 3Academic PressNewYork 1996 p. 93
27	Majoul I Straub M Duden R Hell S W Söling H D Fluorescence resonance energytransfer analysis of protein-protein interactions in single livingcells by multifocal multiphoton microscopyMol. Biotech 2002 82267277

Viewed

Full text

Abstract

Cited

Shared

Discussed