title: Biophysical insights into the interaction of hen egg white lysozyme with therapeutic dye clofazimine: modulation of activity and SDS induced aggregation of model protein.
abstract: The present study details the binding process of clofazimine to hen egg white lysozyme (HEWL) using spectroscopy, dynamic light scattering, transmission electron microscopy (TEM), and molecular docking techniques. Clofazimine binds to the protein with binding constant (Kb) in the order of 1.57 × 104at 298 K. Binding process is spontaneous and exothermic. Molecular docking results suggested the involvement of hydrogen bonding and hydrophobic interactions in the binding process. Bacterial cell lytic activity in the presence of clofazimine increased to more than 40% of the value obtained with HEWL only. Interaction of the drug with HEWL induced ordered secondary structure in the protein and molecular compaction. Clofazimine also effectively inhibited the sodium dodecyl sulfate (SDS) induced amyloid formation in HEWL and caused disaggregation of preformed fibrils, reinforcing the notion that there is involvement of hydrophobic interactions and hydrogen bonding in the binding process of clofazimine with HEWL and clofazimine destabilizes the mature fibrils. Further, TEM images confirmed that fibrillar species were absent in the samples where amyloid induction was performed in the presence of clofazimine. As clofazimine is a drug less explored for the inhibition of fibril formation of the proteins, this study reports the inhibition of SDS-induced amyloid formation of HEWL by clofazimine, which will help in the development of clofazimine-related molecules for the treatment of amyloidosis.
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Mohammad Rehan
authors.initials: MR
authors.lastname: Ajmal
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Sumit Kumar
authors.initials: SK
authors.lastname: Chaturvedi
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Nida
authors.initials: N
authors.lastname: Zaidi
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Parvez
authors.initials: P
authors.lastname: Alam
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Masihuz
authors.initials: M
authors.lastname: Zaman
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Mohammad Khursheed
authors.initials: MK
authors.lastname: Siddiqi
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Saima
authors.initials: S
authors.lastname: Nusrat
authors.affiliation: b King Fahd Medical Research Center , King Abdulaziz University , P.O. Box: 80216, Jeddah 21589 , Saudi Arabia.
authors.forename: Mohammad Sarwar
authors.initials: MS
authors.lastname: Jamal
authors.affiliation: c Deanship of Scientific Research , King Saud University , Riyadh , Saudi Arabia.|d Food Science and Nutrition Department , National Research Center , Dokki, Cairo , Egypt.
authors.forename: Mohamed H
authors.initials: MH
authors.lastname: Mahmoud
authors.affiliation: e Faculty of Science, Zoology Department , Assiut University , Assiut 71516 , Egypt.
authors.forename: Gamal
authors.initials: G
authors.lastname: Badr
authors.affiliation: a Interdisciplinary Biotechnology Unit , Aligarh Muslim University , Aligarh 202002 , India.
authors.forename: Rizwan Hasan
authors.initials: RH
authors.lastname: Khan
mesh_terms: D000682:Amyloid / Q000037:antagonists & inhibitors / Q000737:chemistry*; D000818:Animals; D052160:Benzothiazoles; D001665:Binding Sites; D002645:Chickens; D002991:Clofazimine / Q000737:chemistry*; D004529:Egg White / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D007917:Leprostatic Agents / Q000737:chemistry*; D062105:Molecular Docking Simulation; D009113:Muramidase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D066329:Protein Aggregates; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D054730:Protein Interaction Domains and Motifs; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D013844:Thiazoles / Q000737:chemistry
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D005456:Fluorescent Dyes; D007917:Leprostatic Agents; D066329:Protein Aggregates; D013844:Thiazoles; C009462:thioflavin T; D012967:Sodium Dodecyl Sulfate; D002991:Clofazimine; C421864:hen egg lysozyme; D009113:Muramidase
keywords: Thioflavin T; binding; clofazimine; fluorescence; hen egg white lysozyme