A De novo Peptide from a High Throughput Peptide Library Blocks Myosin A -MTIP Complex Formation in Plasmodium falciparum
Abstract
:1. Introduction
2. Results
2.1. Validation of the Myosin A tail/MTIP Interaction
2.2. Screening Peptide Binder against Myosin A Tail Domain
2.3. ZA1 Interacted with Myosin A Tail In Vitro
2.4. ZA1 Disrupted Myosin A tail-MTIP Complex In Vivo
2.5. ZA1 Disrupted Myosin A tail-MTIP Complex In Vitro
2.6. A Shorter Peptide ZA1S Bound Myosin A Tail and Inhibited the Invasion of P. falciparum
3. Discussion
4. Materials and Methods
4.1. Construction of De Novo Peptide Libraries
4.2. Cloning of Myosin A Tail and MTIP in Bacterial Two-Hybrid Vectors
4.3. Bacterial Two-Hybrid Studies
4.4. Beta Galactosidase Assay
4.5. Cloning, Expression, and Purification of MTIP
4.6. Cloning, Expression, and Purification of ZA1
4.7. ELISA-Based Assay
4.8. Isothermal Calorimetric Assay
4.9. Bacterial Three-Hybrid Studies
4.10. Arabinose Gradient Liquid Beta Galactosidase Assay
4.11. Competitive ELISA
4.12. Competitive Isothermal Calorimetric Assay
4.13. In Silico Studies
4.14. Far Western-Dot Blot Assay
4.15. Parasite Culture
4.16. Ethics Statement
4.17. Growth Inhibition Assay (GIA)
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
MTIP | Myosin A tail interacting protein |
ESAT-6 | Early secreted antigen 6 kDa |
CFP-10 | Culture filtrate protein 10 kDa |
IMC | Inner membrane complex |
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Anam, Z.e.; Joshi, N.; Gupta, S.; Yadav, P.; Chaurasiya, A.; Kahlon, A.K.; Kaushik, S.; Munde, M.; Ranganathan, A.; Singh, S. A De novo Peptide from a High Throughput Peptide Library Blocks Myosin A -MTIP Complex Formation in Plasmodium falciparum. Int. J. Mol. Sci. 2020, 21, 6158. https://doi.org/10.3390/ijms21176158
Anam Ze, Joshi N, Gupta S, Yadav P, Chaurasiya A, Kahlon AK, Kaushik S, Munde M, Ranganathan A, Singh S. A De novo Peptide from a High Throughput Peptide Library Blocks Myosin A -MTIP Complex Formation in Plasmodium falciparum. International Journal of Molecular Sciences. 2020; 21(17):6158. https://doi.org/10.3390/ijms21176158
Chicago/Turabian StyleAnam, Zill e, Nishant Joshi, Sakshi Gupta, Preeti Yadav, Ayushi Chaurasiya, Amandeep Kaur Kahlon, Shikha Kaushik, Manoj Munde, Anand Ranganathan, and Shailja Singh. 2020. "A De novo Peptide from a High Throughput Peptide Library Blocks Myosin A -MTIP Complex Formation in Plasmodium falciparum" International Journal of Molecular Sciences 21, no. 17: 6158. https://doi.org/10.3390/ijms21176158