Abstract
Benzocaine drug (BZC) is an active component of various nonprescription drugs and used for numb teething treatments. The interaction of BZC with bovine serum albumin (BSA) has been studied using fluorescence, synchronous fluorescence, UV-Vis, circular dichroism (CD), and molecular docking analysis. The results revealed that BZC has a strong affinity to quench the intrinsic fluorescence of BSA in terms of a static quenching mechanism under physiological conditions. The fluorescence quenching data revealed that the quenching constants are (KSV) 4.10, 3.30, and 2.35 × 104 L mol−1 at 298, 304, and 310 K, respectively. The binding constants (Kb) at three different temperatures (298, 304, and 310 K) were found to be 6.02, 3.72, and 1.10 × 105 L mol−1, respectively. The thermodynamic parameters ∆H° and ∆S° have been estimated to be − 70.67 and − 128.9 J mol−1 K−1, respectively, thereby, indicating that hydrogen bonding and Van der Waals forces play major role in the interaction of BSA–BZC. Moreover, the negative values of ΔG° − 32.30, − 31.50, − 30.68 kJ mol−1 at 298, 304, 310 K, respectively, indicate the spontaneity of the interaction. FRET analysis proved high probability of energy transfer from BSA to the drug molecule. Molecular docking and displacement studies indicated that BZC was bound to the Sudlow’s site II through hydrogen bonding and Van der Waals interactions.
Similar content being viewed by others
References
Zhang, Q., & Ni, Y. (2017). Comparative studies on the interaction of nitrofuran antibiotics with bovine serum albumin †. https://doi.org/10.1039/c7ra05570f.
Molodenskiy, D., Shirshin, E., Tikhonova, T., Gruzinov, A., Peters, G., & Spinozzi, F. (2017). Thermally induced conformational changes and protein–protein interactions of bovine serum albumin in aqueous solution under different pH and ionic strengths as revealed by SAXS measurements. Physical Chemistry Chemical Physics, 19(26), 17143–17155. https://doi.org/10.1039/C6CP08809K.
Liu, J., He, Y., Liu, D., He, Y., Tang, Z., Lou, H., et al. (2018). Characterizing the binding interaction of astilbin with bovine serum albumin: A spectroscopic study in combination with molecular docking technology. RSC Advances, 8(13), 7280–7286. https://doi.org/10.1039/C7RA13272G.
Ghuman, J., Zunszain, P. A., Petitpas, I., Bhattacharya, A. A., Otagiri, M., & Curry, S. (2005). Structural basis of the drug-binding specificity of human serum albumin. Journal of Molecular Biology, 353(1), 38–52. https://doi.org/10.1016/J.JMB.2005.07.075.
Zhivkova, Z. D. (n.d.). Studies on Drug – Human Serum Albumin Binding: The Current State of the Matter. Retrieved from https://www.ingentaconnect.com/content/ben/cpd/2015/00000021/00000014/art00005
Manea, Y. K., Khan, A. M. T., Qashqoosh, M., Wani, A. A., & Shahadat, M. (2019). Ciprofloxacin-supported chitosan/polyphosphate nanocomposite to bind bovine serum albumin: Its application in drug delivery. Journal of Molecular Liquids, 111337. https://doi.org/10.1016/J.MOLLIQ.2019.111337.
Lou, Y.-Y., Zhou, K.-L., Pan, D.-Q., Shen, J.-L., & Shi, J.-H. (2017). Spectroscopic and molecular docking approaches for investigating conformation and binding characteristics of clonazepam with bovine serum albumin (BSA). Journal of Photochemistry and Photobiology B: Biology, 167, 158–167. https://doi.org/10.1016/J.JPHOTOBIOL.2016.12.029.
Shi, J.-H., Zhou, K.-L., Lou, Y.-Y., & Pan, D.-Q. (2018). Multi-spectroscopic and molecular modeling approaches to elucidate the binding interaction between bovine serum albumin and darunavir, a HIV protease inhibitor. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 188, 362–371. https://doi.org/10.1016/J.SAA.2017.07.040.
Zhou, N., Liang, Y.-Z., & Wang, P. (2007). 18β-Glycyrrhetinic acid interaction with bovine serum albumin. Journal of Photochemistry and Photobiology A: Chemistry, 185(2–3), 271–276. https://doi.org/10.1016/J.JPHOTOCHEM.2006.06.019.
Yang, F., Zhang, Y., Liang, H., Yang, F., Zhang, Y., & Liang, H. (2014). Interactive Association of Drugs Binding to human serum albumin. International Journal of Molecular Sciences, 15(3), 3580–3595. https://doi.org/10.3390/ijms15033580.
Mangiapia, G., Gvaramia, M., Kuhrts, L., Teixeira, J., Koutsioubas, A., Soltwedel, O., & Frielinghaus, H. (2017). Effect of benzocaine and propranolol on phospholipid-based bilayers. Physical Chemistry Chemical Physics, 19(47), 32057–32071. https://doi.org/10.1039/C7CP06077G.
Mura, P., Maestrelli, F., González-Rodríguez, M. L., Michelacci, I., Ghelardini, C., & Rabasco, A. M. (2007). Development, characterization and in vivo evaluation of benzocaine-loaded liposomes. European Journal of Pharmaceutics and Biopharmaceutics, 67(1), 86–95. https://doi.org/10.1016/J.EJPB.2007.01.020.
Li, S., & Li, D. (2011). Investigation on the pH-dependent binding of benzocaine and lysozyme by fluorescence and absorbance. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 82(1), 396–405. https://doi.org/10.1016/J.SAA.2011.07.069.
Zhang, H., Mei, P., & Yang, X. (2009). Optical, structural and thermodynamic properties of the interaction between tradimefon and serum albumin. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 72(3), 621–626. https://doi.org/10.1016/J.SAA.2008.10.062.
Chinnathambi, S., Velmurugan, D., Hanagata, N., Aruna, P. R., & Ganesan, S. (2014). Investigations on the interactions of 5-fluorouracil with bovine serum albumin: Optical spectroscopic and molecular modeling studies. Journal of Luminescence, 151, 1–10. https://doi.org/10.1016/J.JLUMIN.2014.01.063.
Silva, D., Cortez, C. M., Cunha-Bastos, J., & Louro, S. R. (2004). Methyl parathion interaction with human and bovine serum albumin. Toxicology Letters, 147(1), 53–61. https://doi.org/10.1016/J.TOXLET.2003.10.014.
Ghisaidoobe, A. B. T., & Chung, S. J. (2014). Intrinsic tryptophan fluorescence in the detection and analysis of proteins: A focus on Förster resonance energy transfer techniques. International Journal of Molecular Sciences, 15(12), 22518–22538. https://doi.org/10.3390/ijms151222518.
Zhang, G., Que, Q., Pan, J., & Guo, J. (2008). Study of the interaction between icariin and human serum albumin by fluorescence spectroscopy. Journal of Molecular Structure, 881(1–3), 132–138. https://doi.org/10.1016/J.MOLSTRUC.2007.09.002.
Shen, H., Gu, Z., Jian, K., & Qi, J. (2013). In vitro study on the binding of gemcitabine to bovine serum albumin. Journal of Pharmaceutical and Biomedical Analysis, 75, 86–93. https://doi.org/10.1016/J.JPBA.2012.11.021.
Jana, S., Dalapati, S., Ghosh, S., & Guchhait, N. (2012). Binding interaction between plasma protein bovine serum albumin and flexible charge transfer fluorophore: a spectroscopic study in combination with molecular docking and molecular dynamics simulation. Journal of Photochemistry and Photobiology A: Chemistry, 231(1), 19–27. https://doi.org/10.1016/J.JPHOTOCHEM.2011.12.002.
Akdogan, Y., Emrullahoglu, M., Tatlidil, D., Ucuncu, M., & Cakan-Akdogan, G. (2016). EPR studies of intermolecular interactions and competitive binding of drugs in a drug–BSA binding model. Physical Chemistry Chemical Physics, 18(32), 22531–22539. https://doi.org/10.1039/C6CP04137J.
Shaikh, S. M. T., Seetharamappa, J., Ashoka, S., & Kandagal, P. B. (2006). Spectroscopic studies and life time measurements of binding of a bioactive compound to bovine serum albumin and the effects of common ions and other drugs on binding. Chemical & Pharmaceutical Bulletin, 54(4), 422–427. https://doi.org/10.1248/cpb.54.422.
Peng, X., Qi, W., Huang, R., Su, R., & He, Z. (2015). Elucidating the influence of gold nanoparticles on the binding of salvianolic acid B and rosmarinic acid to bovine serum albumin. PLoS One, 10(4), e0118274. https://doi.org/10.1371/journal.pone.0118274.
Hu, Y.-J., Liu, Y., Zhao, R.-M., Dong, J.-X., & Qu, S.-S. (2006). Spectroscopic studies on the interaction between methylene blue and bovine serum albumin. Journal of Photochemistry and Photobiology A: Chemistry, 179(3), 324–329. https://doi.org/10.1016/J.JPHOTOCHEM.2005.08.037.
Becker, D. E., & Reed, K. L. (2012). Local anesthetics: review of pharmacological considerations. Anesthesia Progress, 59(2), 90–101; quiz 102–3. https://doi.org/10.2344/0003-3006-59.2.90.
Lakowicz, J. R., & Masters, B. R. (2008). Principles of fluorescence spectroscopy, third edition. Journal of Biomedical Optics, 13(2), 029901. https://doi.org/10.1117/1.2904580.
Athina Papadopoulou, Rebecca J. Green, , & Richard A. Frazier. (2004). Interaction of flavonoids with bovine serum albumin: a fluorescence quenching study. doi:https://doi.org/10.1021/JF048693G.
Zhang, G., Wang, A., Jiang, T., & Guo, J. (2008). Interaction of the irisflorentin with bovine serum albumin: a fluorescence quenching study. Journal of Molecular Structure, 891(1–3), 93–97. https://doi.org/10.1016/J.MOLSTRUC.2008.03.002.
Khan, A. B., Khan, J. M., Ali, M. S., Khan, R. H., & Kabir-ud-Din. (2012). Interaction of amphiphilic drugs with human and bovine serum albumins. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 97, 119–124. https://doi.org/10.1016/J.SAA.2012.05.060.
Amaral, M., Kokh, D. B., Bomke, J., Wegener, A., Buchstaller, H. P., Eggenweiler, H. M., et al. (2017). Protein conformational flexibility modulates kinetics and thermodynamics of drug binding. Nature Communications, 8(1), 2276. https://doi.org/10.1038/s41467-017-02258-w.
Ross, P. D., & Subramanian, S. (1981). Thermodynamics of protein association reactions: forces contributing to stability. Biochemistry, 20(11), 3096–3102. https://doi.org/10.1021/bi00514a017.
Yasseen, Z., & Omar El-Ghossain, M. (2016). Studies on binding of widely used drugs with human serum albumin at different temperatures and PHs radiation measurements in the environment view project a Flureescence quenching studies of drugs binding with human serum albumin view project. https://doi.org/10.4172/2254-609X.100033.
Zhao, X., Liu, R., Teng, Y., & Liu, X. (2011). The interaction between ag+ and bovine serum albumin: A spectroscopic investigation. Science of the Total Environment, 409(5), 892–897. https://doi.org/10.1016/J.SCITOTENV.2010.11.004.
Samari, F., Hemmateenejad, B., Rezaei, Z., & Shamsipur, M. (2012). A novel approach for rapid determination of vitamin B12 in pharmaceutical preparations using BSA-modified gold nanoclusters. Analytical Methods, 4(12), 4155. https://doi.org/10.1039/c2ay25196e.
Ahmad, B., Parveen, S., & Khan, R. H. (2006). Effect of albumin conformation on the binding of ciprofloxacin to human serum albumin: a novel approach directly assigning binding site. https://doi.org/10.1021/BM050996B.
Liu, X.-H., Xi, P.-X., Chen, F.-J., Xu, Z.-H., & Zeng, Z.-Z. (2008). Spectroscopic studies on binding of 1-phenyl-3-(coumarin-6-yl)sulfonylurea to bovine serum albumin. Journal of Photochemistry and Photobiology B: Biology, 92(2), 98–102. https://doi.org/10.1016/J.JPHOTOBIOL.2008.04.008.
Hu, Y.-J., Liu, Y., Shen, X.-S., Fang, X.-Y., & Qu, S.-S. (2005). Studies on the interaction between 1-hexylcarbamoyl-5-fluorouracil and bovine serum albumin. Journal of Molecular Structure, 738(1–3), 143–147. doi:https://doi.org/10.1016/J.MOLSTRUC.2004.11.062.
Sudlow, G., Birkett, D. J., & Wade, D. N. (1976). Further characterization of specific drug binding sites on human serum albumin. Molecular Pharmacology, 12(6).
Yamasaki, K., Maruyama, T., Kragh-Hansen, U., & Otagiri, M. (1996). Characterization of site I on human serum albumin: concept about the structure of a drug binding site. Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1295(2), 147–157. https://doi.org/10.1016/0167-4838(96)00013-1.
Huang, B. X., Kim, H.-Y., & Dass, C. (2004). Probing three-dimensional structure of bovine serum albumin by chemical cross-linking and mass spectrometry. Journal of the American Society for Mass Spectrometry, 15(8), 1237–1247. https://doi.org/10.1016/j.jasms.2004.05.004.
Cheng, Z., & Zhang, Y. (2008). Fluorometric investigation on the interaction of oleanolic acid with bovine serum albumin. Journal of Molecular Structure, 879(1–3), 81–87. https://doi.org/10.1016/J.MOLSTRUC.2007.08.020.
Acknowledgments
The authors are grateful to the Interdisciplinary Biotechnology Unit, Aligarh Muslim University, India for supporting this work.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest.
Research Involving Humans and Animals Statement
None.
Informed Consent
None.
Funding Information
One of the authors Mohsen Qashqoosh received financial assistance from University Grant Commission, India.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Qashqoosh, M.T.A., Manea, Y.K., Alahdal, F.A.M. et al. Investigation of Conformational Changes of Bovine Serum Albumin upon Binding with Benzocaine Drug: a Spectral and Computational Analysis. BioNanoSci. 9, 848–858 (2019). https://doi.org/10.1007/s12668-019-00663-7
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12668-019-00663-7