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A spectroscopic and computational intervention of interaction of lysozyme with 6-mercaptopurine


In the present work, biophysical insight into the binding interactions of the protein, hen egg white (HEW) lysozyme (Lyz) with an anticancer drug, 6-mercaptopurine (6-MP)‚ was investigated by using a combination of spectroscopic and computational tools. 6-MP, a synthetic analog of natural purines, is a well-known anticancer drug and antiviral agent that inhibits the synthesis of RNA, DNA, and proteins. Lysozyme is a single-chain protein that can combine with endogenous and exogenous substances to exert its antiviral, antibacterial, and antitumor effects. The intrinsic fluorescence of lysozyme was quenched with the increased addition of 6-MP. The quenching mechanism was found to be static in nature as shown by the fluorescence lifetime and excitation spectrum measurements. The conformational changes of Lyz in the presence of 6-MP were monitored both at the ensemble and single-molecule level by using synchronous fluorescence spectroscopy, circular dichroism (CD), and fluorescence correlation spectroscopy (FCS). Molecular docking results predicted the probable binding sites for 6-MP on Lyz. The experimental findings are in good agreement with the results obtained by the molecular dynamics (MD) simulation study.

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The authors thank Mr. Ajaya Das, Chemical Sciences Division, Saha Institute of Nuclear Physics (SINP), Kolkata, India for his assistance in time-resolved fluorescence measurements and preliminary data analysis. We also thank Dr. Anand Kant Das and Dr. Shrutidhara Biswas for the helpful scientific discussion.


The authors acknowledge the Indian Institute of Technology (Indian School of Mines), Dhanbad, India for infrastructure facility and financial assistance (FRS (74)/2014-15/APH). For the computational facility, we are thankful to the Ministry of Human Resource Development (MHRD), New Delhi, India for financial assistance (MHRD/(FDC)/2015-2016/438/INST).

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Correspondence to Umakanta Tripathy.

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Swain, B.C., Mukherjee, S.K., Rout, J. et al. A spectroscopic and computational intervention of interaction of lysozyme with 6-mercaptopurine. Anal Bioanal Chem (2020).

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  • Lysozyme
  • 6-Mercaptopurine
  • Fluorescence spectroscopy
  • Circular dichroism
  • Fluorescence correlation spectroscopy
  • MD simulation