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Spectroscopic analysis of the interaction between bilirubin and bovine serum albumin

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Abstract.

The interactions between bilirubin (BR) and bovine serum albumin (BSA) have been studied by fluorescence spectroscopy. The association constant between BR and BSA was obtained by fluorescence enhancement titration. Furthermore, fluorescence quenching was studied at different temperatures, and the binding constant was also determined by the method of fluorescence quenching. The two methods yielded similar results. It indicated that the former method could be successfully applied to the determination of BR. The results showed that the binding of BR to BSA induced conformational changes in BSA. Based on the theory of Förster energy transfer, the distance between BR and protein were calculated. According to the thermodynamic parameters, the main binding force could be judged. The experimental results revealed that BSA and BR had strong interactions. The mechanism of quenching belonged to static quenching and the main sort of binding force was van der Waals interactions and hydrogen bonds.

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References

  • S F Asad S Singh A Ahmad N U Khan S M Hadi (2001) ArticleTitleProoxidant and antioxidant activities of bilirubin and its metabolic precursor biliverdin: a structure-activity study J Chem-Biol Interact 137 59 Occurrence Handle10.1016/S0009-2797(01)00209-5 Occurrence Handle1:CAS:528:DC%2BD3MXmtV2nurw%3D

    Article  CAS  Google Scholar 

  • R Stocker A N Glazer B N Ames (1987) ArticleTitleAntioxidant activity of albumin-bound bilirubin Proc Natl Acad Sci USA 84 5918 Occurrence Handle10.1073/pnas.84.16.5918 Occurrence Handle1:CAS:528:DyaL1cXltFKmsg%3D%3D

    Article  CAS  Google Scholar 

  • M Minetti C Mallozzi A M M D Stasi D Pietraforte (1998) ArticleTitleBilirubin is an effective antioxidant of peroxynitrite-mediated protein oxidation in human blood plasma J Biochem Biophys 352 165 Occurrence Handle10.1006/abbi.1998.0584 Occurrence Handle1:CAS:528:DyaK1cXislahs78%3D

    Article  CAS  Google Scholar 

  • T W R Hansen (2000) ArticleTitleBilirubin oxidation in brain J Mol Genet Metab 71 411 Occurrence Handle10.1006/mgme.2000.3028 Occurrence Handle1:CAS:528:DC%2BD3cXms1yjsb4%3D

    Article  CAS  Google Scholar 

  • S M Shapiro (2003) ArticleTitleBilirubin toxicity in the developing nervous system J Pediatr Neurol 29 410 Occurrence Handle10.1016/j.pediatrneurol.2003.09.011

    Article  Google Scholar 

  • M A Brito D Brites D A Butterfield (2004) ArticleTitleA link between hyperbilirubinemia, oxidative stress and injury to neocortical synaptosomes Brain Res 1026 33 Occurrence Handle10.1016/j.brainres.2004.07.063 Occurrence Handle1:CAS:528:DC%2BD2cXot12jtLc%3D

    Article  CAS  Google Scholar 

  • D C Carter J X Ho (1994) ArticleTitleStructure of serum albumin Adv Protein Chem 45 153 Occurrence Handle1:CAS:528:DyaK2cXltVajtLk%3D

    CAS  Google Scholar 

  • R D Gray S D Stroupe (1978) ArticleTitleKinetics and mechanism of bilirubin binding to human serum albumin J Biol Chem 253 4370 Occurrence Handle1:CAS:528:DyaE1cXkslahtr8%3D

    CAS  Google Scholar 

  • F L Cui J Fan J P Li Z D Hu (2004) ArticleTitleInteractions between 1-benzoyl-4-p-chlorophenyl thio-semicarbazide and serum albumin: investigation by fluorescence spectroscopy Bioorg Med Chem 12 151 Occurrence Handle10.1016/j.bmc.2003.10.018 Occurrence Handle1:CAS:528:DC%2BD3sXhtVSjtLfN

    Article  CAS  Google Scholar 

  • R Brodersen (1974) ArticleTitleCompetitive binding of bilirubin and drugs to human serum albumin studied by enzymatic oxidation J Clin Invest 54 1353 Occurrence Handle1:CAS:528:DyaE2MXpt12rsQ%3D%3D Occurrence Handle10.1172/JCI107882

    Article  CAS  Google Scholar 

  • M A Khan S Muzammil J Musarrat (2002) ArticleTitleDifferential binding of tetracyclines with serum albumin and induced structural alterations in drug-bound protein J Biol Macromol 30 243 Occurrence Handle10.1016/S0141-8130(02)00038-7 Occurrence Handle1:CAS:528:DC%2BD38Xnt1Giu7c%3D

    Article  CAS  Google Scholar 

  • J H Tang F Luan X G Chen (2006) ArticleTitleBinding analysis of glycyrrhetinic acid to human serum albumin: Fluorescence spectroscopy, FTIR, and molecular modeling Bioorg Med Chem 14 3210 Occurrence Handle10.1016/j.bmc.2005.12.034 Occurrence Handle1:CAS:528:DC%2BD28Xis1ehs7c%3D

    Article  CAS  Google Scholar 

  • P B Kandagal S Ashoka J Seetharamappa V Vani S M T Shaikh (2006) ArticleTitleStudy of the interaction between doxepin and human serum albumin by spectroscopic methods J Photochem Photobio A 179 161 Occurrence Handle10.1016/j.jphotochem.2005.08.008 Occurrence Handle1:CAS:528:DC%2BD28Xit1ajsbw%3D

    Article  CAS  Google Scholar 

  • S Tayyab N J Khan M A Khan Y Kumar (2003) ArticleTitleBehavior of various mammalian albumins towards bilirubin binding and photochemical properties of different bilirubin-albumin complexes Int J Biol Macromol 31 187 Occurrence Handle10.1016/S0141-8130(02)00081-8 Occurrence Handle1:CAS:528:DC%2BD3sXptFCksg%3D%3D

    Article  CAS  Google Scholar 

  • M M Khan S Tayyab (2001) ArticleTitleUnderstanding the role of internal lysine residues of serum albumins in conformational stability and bilirubin binding Biochim Biophys Acta 1545 263 Occurrence Handle1:CAS:528:DC%2BD3MXpsFKktw%3D%3D

    CAS  Google Scholar 

  • B I Greene A A Lamola C V Shank (1981) ArticleTitlePicosecond primary photoprocesses of bilirubin bound to human serum albumin Proc Natl Acad Sci USA 78 2008 Occurrence Handle10.1073/pnas.78.4.2008 Occurrence Handle1:CAS:528:DyaL3MXktlOltL0%3D

    Article  CAS  Google Scholar 

  • S Tayyab N Sharma M M Khan (2000) ArticleTitleUse of domain specific ligands to study urea-induced unfolding of bovine serum albumin Biochem Biophys Res Commun 277 83 Occurrence Handle10.1006/bbrc.2000.3642 Occurrence Handle1:CAS:528:DC%2BD3cXntFOqurw%3D

    Article  CAS  Google Scholar 

  • L T Lemiesz A Karaczyn B K Keppler H Kozlowski (2000) ArticleTitleStudies on the interactions between human serum albumin and trans-indazolium (bisindazole) tetrachlororuthenate(III) J Inorg Biochem 78 341 Occurrence Handle10.1016/S0162-0134(00)00062-3

    Article  Google Scholar 

  • J Jacobsen (1977) ArticleTitleStudies of the affinity of human serum albumin for binding of bilirubin at different temperatures and ionic strength Int J Pept Protein Res 9 235 Occurrence Handle1:CAS:528:DyaE2sXhtFKhtLs%3D Occurrence Handle10.1111/j.1399-3011.1977.tb03486.x

    Article  CAS  Google Scholar 

  • C B Berde B S Hudson R D Simoni L A Sklar (1979) ArticleTitleHuman serum albumin: Spectroscopic studies of binding and proximity relationships for fatty acids and bilirubin J Biol Chem 254 391 Occurrence Handle1:CAS:528:DyaE1MXpvVyktg%3D%3D

    CAS  Google Scholar 

  • R Brodersen (1979) ArticleTitleBilirubin: Solubility and interaction with albumin and phospholipid J Biol Chem 254 2364 Occurrence Handle1:CAS:528:DyaE1MXhvF2iu74%3D

    CAS  Google Scholar 

  • B Zhang Y Fung K Lau B Lin (1999) ArticleTitleBilirubin-human serum albumin interaction monitored by capillary zone electrophoresis Biomed Chromatogr 13 267 Occurrence Handle10.1002/(SICI)1099-0801(199906)13:4<267::AID-BMC833>3.0.CO;2-L Occurrence Handle1:CAS:528:DyaK1MXksFOlsr8%3D

    Article  CAS  Google Scholar 

  • H Athar N Ahmad S Tayyab M A Qasim (1999) ArticleTitleUse of fluorescence enhancement technique to study bilirubin-albumin interaction J Biol Macromol 25 353 Occurrence Handle10.1016/S0141-8130(99)00056-2 Occurrence Handle1:CAS:528:DyaK1MXktlygs7w%3D

    Article  CAS  Google Scholar 

  • R L Levine (1977) ArticleTitleFluorescence-quenching studies of the binding of bilirubin to albumin J Clin Chem 23 2292 Occurrence Handle1:CAS:528:DyaE1cXjsVWjsw%3D%3D

    CAS  Google Scholar 

  • X P Li Z Rosenzweig (1997) ArticleTitleA fiber optic sensor for rapid analysis of bilirubin in serum Anal Chimica Acta 353 263 Occurrence Handle10.1016/S0003-2670(97)87785-9 Occurrence Handle1:CAS:528:DyaK2sXnvVShtbw%3D

    Article  CAS  Google Scholar 

  • J T Coleman J F Eastham M J Sepaniak (1984) ArticleTitleFiber optic based sensor for bioanalytical absorbance measurements Anal Chem 56 2246 Occurrence Handle10.1021/ac00276a060 Occurrence Handle1:CAS:528:DyaL2cXlt1KjtbY%3D

    Article  CAS  Google Scholar 

  • G Z Chen X Z Huang J G Xu Z Z Zheng Z B Wang (1990) The methods of fluorescence analysis EditionNumber2 Science Press Beijing 112

    Google Scholar 

  • Y Xu H X Shen H G Huan (1997) ArticleTitleFluorescence characteristics of iron(III)-norfloxacin-guanylic acid ternary complex Anal Chem 25 419 Occurrence Handle1:CAS:528:DyaK2sXjtVChtbc%3D

    CAS  Google Scholar 

  • R M Clegg (1995) ArticleTitleFluorescence resonance energy transfer Curr Opin Biotechnol 6 103 Occurrence Handle10.1016/0958-1669(95)80016-6 Occurrence Handle1:CAS:528:DyaK2MXjsl2ks7Y%3D

    Article  CAS  Google Scholar 

  • Y J Hu Y Liu R M Zhao J X Dong S Sh Qu (2006) ArticleTitleSpectroscopic studies on the interaction between methylene blue and bovine serum albumin J Photochem Photobio A 179 324 Occurrence Handle10.1016/j.jphotochem.2005.08.037 Occurrence Handle1:CAS:528:DC%2BD28XisFCjsrY%3D

    Article  CAS  Google Scholar 

  • C Q Jiang M X Gao J X He (2002) ArticleTitleStudy of the interaction between terazosin and serum albumin: synchronous fluorescence determination of terazoain Anal Chim Acta 452 185 Occurrence Handle10.1016/S0003-2670(01)01453-2 Occurrence Handle1:CAS:528:DC%2BD38Xmtlaguw%3D%3D

    Article  CAS  Google Scholar 

  • P D Ross S Subramanian (1981) ArticleTitleThermodynamics of protein association reactions: forces contributing to stability Biochemistry 20 3096 Occurrence Handle10.1021/bi00514a017 Occurrence Handle1:CAS:528:DyaL3MXktF2qsb0%3D

    Article  CAS  Google Scholar 

  • Y H Pang L Yang S M Shuang C Dong M Thompson (2005) ArticleTitleInteraction of human serum albumin with bendroflumethiazide studied by fluorescence spectroscopy J Photochem Photobio B 80 139 Occurrence Handle10.1016/j.jphotobiol.2005.03.006 Occurrence Handle1:CAS:528:DC%2BD2MXmsVWksr4%3D

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Ministry-of-Education Key Laboratory for the Synthesis and Application of Organic Functional Molecules, Hubei University, 430062, Wuhan City, Hubei Province, P.R. China

    Juan Chen, Gongwu Song, Yu He & Qiujun Yan

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  1. Juan Chen
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  2. Gongwu Song
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  3. Yu He
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  4. Qiujun Yan
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Correspondence to Gongwu Song.

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Chen, J., Song, G., He, Y. et al. Spectroscopic analysis of the interaction between bilirubin and bovine serum albumin. Microchim Acta 159, 79–85 (2007). https://doi.org/10.1007/s00604-007-0736-9

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  • DOI: https://doi.org/10.1007/s00604-007-0736-9

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