| Autor: |
Doharey PK; Department of Biochemistry, University of Allahabad, Allahabad, India., Verma P; Biochemistry Division, CSIR-Central Drug Research Institute, Lucknow, India., Dubey A; Computational Chemistry and Drug discovery Division, Quanta calculus Pvt. Ltd, Kushinagar, India.; Department of Pharmacology, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences, Chennai, India., Singh SK; Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India., Kumar M; Department of Biochemistry, University of Allahabad, Allahabad, India., Tripathi T; Department of Biochemistry, North-Eastern Hill University, Umshing, India., Alonazi M; Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia., Siddiqi NJ; Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia., Sharma B; Department of Biochemistry, University of Allahabad, Allahabad, India. |
| Abstrakt: |
Human Lymphatic filariasis is caused by parasitic nematodes Wuchereria bancrofti, Brugia malayi, and Brugia timori. Protein disulfide isomerase (PDI), a redox-active enzyme, helps to form and isomerize the disulfide bonds, thereby acting as a chaperone. Such activity is essential for activating many essential enzymes and functional proteins. Brugia malayi protein disulfide isomerase (BmPDI) is crucial for parasite survival and an important drug target. Here, we used a combination of spectroscopic and computational analysis to study the structural and functional changes in the BmPDI during unfolding. Tryptophan fluorescence data revealed two well-separated transitions during the unfolding process, suggesting that the unfolding of the BmPDI is non-cooperative. The binding of the fluorescence probe 8-anilino-1-naphthalene sulfonic acid dye (ANS) validated the results obtained by the pH unfolding. The dynamics of molecular simulation performed at different pH conditions revealed the structural basis of BmPDI unfolding. Detailed analysis suggested that under different pH, both the global structure and the conformational dynamics of the active site residues were differentially altered. Our multiparametric study reveals the differential dynamics and collective motions of BmPDI unfolding, providing insights into its structure-function relationship.Communicated by Ramaswamy H. Sarma. |
