Molecular cloning and characterization of protein disulfide isomerase of Brugia malayi, a human lymphatic filarial parasite

Authors

  • Pravesh Verma Division of Biochemistry, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India
  • Pawan Kumar Doharey Division of Biochemistry, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India
  • Sunita Yadav Division of Biochemistry, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India
  • Ankur Omer Division of Toxicology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India
  • Poonam Singh Division of Toxicology, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India
  • Jitendra Kumar Saxena Division of Biochemistry, CSIR-Central Drug Research Institute, BS10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India

DOI:

https://doi.org/10.17179/excli2017-214

Keywords:

protein disulfide isomerase, Brugia malayi, antifilarials, homology modelling and docking

Abstract

Lymphatic filariasis results in an altered lymphatic system and the abnormal enlargement of body parts, causing pain, serious disability and social stigma. Effective vaccines are still not available nowadays, drugs against the disease is required. Protein disulfide isomerase (PDI) is an essential catalyst of the endoplasmic reticulum which is involved in folding and chaperone activities in different biological systems. Here, we report the enzymatic characterization of a Brugia malayi Protein disulfide isomerase (BmPDI), which was expressed and purified from Escherichia coli BL21 (DE3). Western blotting analysis showed the recombinant BmPDI could be recognized by anti-BmPDI Rabbit serum. The rBmPDI exhibited an optimum activity at pH 8 and 40 °C. The enzyme was inhibited by aurin and PDI inhibitor. Recombinant BmPDI showed interaction with recombinant Brugia malayi calreticulin (rBmCRT). The three-dimensional model for BmPDI and BmCRT was generated by homology modelling. A total of 25 hydrogen bonds were found to be formed between two interfaces. There are 259 non-bonded contacts present in the BmPDI-BmCRT complex and 12 salt bridges were formed in the interaction.

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Published

2017-06-01

How to Cite

Verma, P., Doharey, P. K., Yadav, S., Omer, A., Singh, P., & Saxena, J. K. (2017). Molecular cloning and characterization of protein disulfide isomerase of Brugia malayi, a human lymphatic filarial parasite. EXCLI Journal, 16, 824–839. https://doi.org/10.17179/excli2017-214

Issue

Vol. 16 (2017)

Section

Original articles

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