Kumar, Krishan and Banerjee, S and Sanivarapu, H and Veronica, J and Namdeo, M and Anand, A and Sundar, S and Swamy, M J and Maurya, R (2024) Characterization of differentially regulated carboxypeptidase (metallopeptidase M32) protein in Miltefosine resistant Leishmania donovani parasites. International Journal of Biological Macromolecules (TSI), 289. ISSN 0141-8130
Full text not available from this repository. (Request a copy)Abstract
Introduction Carboxypeptidase, a member of the metallopeptidase M32 family, catalyses the C-terminal hydrolysis of a variety of peptides and proteins in the presence of metal ions. Objective To characterize Leishmania donovani carboxypeptidase (LdCP) in miltefosine (MIL) drug-resistant parasites. Methods We performed the MTT assay and cell cycle analysis to confirm the MIL resistance of clinical isolates. LdCP gene was cloned and expressed in E. coli Artic strain. The purified LdCP protein was used for antibody generation and biochemical characterization. Results MTT assay and cell cycle analysis revealed that all three isolates exhibit MIL resistance. LdCP constitutively expressed in both promastigote and amastigote stages of parasites, and its activity increased 2–3 fold in MIL-resistant parasites. LdCP has high α-helical content at physiological pH and temperature. The protein is quite thermostable with a Tm of 63 °C and susceptible to chemical denaturation, with 50 % unfolding induced by 3.59 M urea or 0.31 M guanidine hydrochloride (GdmCl). LC-MS/MS study reveals that LdCP interacts with membrane-associated proteins that have ATP binding sites and involved in protein phosphorylation. Conclusion To our knowledge, this is the first study to characterize the carboxypeptidase of L. donovani that appears to contribute to the development of MIL resistance parasites.
| Item Type: | Article |
|---|---|
| Divisions: | Global Research Program - Accelerated Crop Improvement |
| CRP: | UNSPECIFIED |
| Uncontrolled Keywords: | carboxypeptidase, Miltefosine resistant, Leishmania donovani parasites |
| Subjects: | Others > Plant Pathology |
| Depositing User: | Mr Nagaraju T |
| Date Deposited: | 20 Mar 2025 04:04 |
| Last Modified: | 20 Mar 2025 04:04 |
| URI: | http://oar.icrisat.org/id/eprint/13004 |
| Official URL: | https://www.sciencedirect.com/science/article/abs/... |
| Projects: | UNSPECIFIED |
| Funders: | UNSPECIFIED |
| Acknowledgement: | This work was supported by Research Grant No. 6/9-7(311) 2023-ECD-II from the Indian Council of Medial Research (ICMR), India to RM. MJS acknowledges Grant No. UOH-IOE-RC3-21-015 from the Institution of Eminence project of University of Hyderabad sanctioned by UGC (India). We extend our sincere thanks to the Instrumentation facility of School of Life sciences, University of Hyderabad for their help and support for providing access to Confocal microscopy and CD spectroscopy. We are also thankful.... |
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