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Isolation and Characterization of Heat Shock Protein 70-Ipek 1 from Toxoplasma gondii

Year 2024, , 496 - 502, 30.09.2024

Kübra Açıkalın Coşkun , Lütfi Tutar

https://doi.org/10.17776/csj.1501849

Abstract

Toxoplasma gondii is a common intracellular parasite that causes the toxoplasmosis. Heat shock proteins (Hsps) have a critical role in pathogenesis of toxoplasmosis. Hsps are highly conserved proteins in evolution among living organisms. This protein family responsible for a wide range of biological processes such as protein folding, protein translocation, protein aggregation. In the present study, Hsp70, a member of the Hsp family, was isolated from T. gondii and its sequence and motifs were determined by PCR, cloning, sequencing and homology modelling analysis. ATP hydrolysis, luciferase folding, and luciferase aggregation experiments were performed for determination of its chaperone activity while the stability and secondary structure of the Hsp70 were discovered by using biophysical experiments (FTIR, florescence and quenching experiment). In addition, in silico analysis were used to determine the physicochemical characteristics of Hsp70. The results revealed that Hsp70 protein obtained from T. gondii (Hsp70-IPEK1) is similar to Hsp70s from other organisms. Also, the chaperone activity, stability and secondary structure of Hsp70-IPEK1 were determined. Hsp70-IPEK1 together with other chaperones in the presence of nucleotide were dramatically increased protein folding and aggregation. According to these results, it is thought that Hsp70 has a potential to contribute many research areas such as pharmaceutical analysis

Keywords

Toxoplasma gondii Heat Shock Protein 70 Stress

References

  • [1] Coşkun K.A., Tutar Y., Isolation and characterization of Heat Shock Protein 100-Batu1 from Toxoplasma gondii RH strain, Exp. Parasitol, 153 (2015) 91–97.
  • [2] Sun H., Zhuo X., Zhao X., et al., The heat shock protein 90 of Toxoplasma gondii is essential for invasion of host cells and tachyzoite growth, Parasite, 24 (2017) 1–11.
  • [3] Ashwinder K., Kho M.T., Chee P.M., et al., Targeting Heat Shock Proteins 60 and 70 of Toxoplasma gondii as a Potential Drug Target: In Silico Approach, Interdiscip. Sci. Comput. Life Sci., 8 (2015) 374–387.
  • [4] de Barros, R. A. M., Torrecilhas, A. C., Marciano, M. A. M., Mazuz, M. L., Pereira-Chioccola, V. L., & Fux, B. Toxoplasmosis in human and animals around the world. Diagnosis and perspectives in the one health approach. Acta tropica, (2022) 231, 106432.
  • [5] Angel, S. O., Vanagas, L., & Alonso, A. M. (2024). Mechanisms of adaptation and evolution in Toxoplasma gondii, Molecular and Biochemical Parasitology, (2024), 258, 111615.
  • [6] Tutar L., Tutar Y., Heat Shock Proteins; An Overview, Curr. Pharm. Biotechnol., 11 (2010) 216–222.
  • [7] Sherman M.Y., Gabai V.L., Hsp70 in cancer: Back to the future, Oncogene, (2014) 1–9.
  • [8] Tran N.T., Jakovlic I., Wang W.M., In silico characterisation, homology modelling and structure-based functional annotation of blunt snout bream (Megalobrama amblycephala) Hsp70 and Hsc70 proteins, J. Anim. Sci. Technol., 57 (2015) 1–9.
  • [9] Sharma D., Masison D.C., Hsp70 structure, function, regulation and influence on yeast prions, Protein Pept. Lett., 16 (2009) 571–81.
  • [10] Tutar Y., Okan Ş., Heat shock protein 70 purification and characterization from Cyprinion macrastomus and Garra rufa obtusa, J. Therm. Biol., 37 (2012) 95–99.
  • [11] Guruprasad K., Reddy B.V.B., Pandit M.W., Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence, Protein Eng. Des. Sel., 4 (1990) 155–161.
  • [12] Islam M.S., Shahik S.M., Sohel M., et al., In Silico Structural and Functional Annotation of Hypothetical Proteins of Vibrio cholerae O139, Genomics Inform., 13 (2015) 53–59.
  • [13] Evans C.G., Chang L., Gestwick J.E., Heat Shock Protein 70 (Hsp70) as an Emerging Drug Target, J. Med. Chem., 53 (2010) 4585–4602.
  • [14] Tutar L., Is Hsp70 a folder or a modulator protein?, Basic Clin. Sci., (2010) 18–24.
  • [15] Pallavi R., Roy N., Nageshan R.K., et al., Heat Shock Protein 90 as a Drug Target against Protozoan Infections, J. Biol. Chem., 285 (2010) 37964–37975.
  • [16] Mammari N., Halabi, Mohamad Adnan Yaacoub S., Chlala H., Dardé, Marie-Laure Courtioux B., Toxoplasma gondii Modulates the Host Cell Responses: An Overview of Apoptosis Pathways, Biomed. Res. Int., (2019) 1–10.
  • [17] Metzger D.C.H., Hemmer-Hansen J., Schulte P.M., Conserved structure and expression of Hsp70 paralogs in teleost fishes, Comp. Biochem. Physiol. Part D Genomics Proteomics, 18 (2016) 10–20.
  • [18] Luengo T.M., Mayer M.P., Rüdiger S.G.D., The Hsp70 – Hsp90 Chaperone Cascade in Protein Folding, Trends Cell Biol., (2018) 1–14.
  • [19] Mayer M.P., Bukau B., Hsp70 chaperones : Cellular functions and molecular mechanism, Cell Mol. Life Sci., 62 (2005) 670–684.

Year 2024, , 496 - 502, 30.09.2024

Kübra Açıkalın Coşkun , Lütfi Tutar

https://doi.org/10.17776/csj.1501849

Abstract

References

  • [1] Coşkun K.A., Tutar Y., Isolation and characterization of Heat Shock Protein 100-Batu1 from Toxoplasma gondii RH strain, Exp. Parasitol, 153 (2015) 91–97.
  • [2] Sun H., Zhuo X., Zhao X., et al., The heat shock protein 90 of Toxoplasma gondii is essential for invasion of host cells and tachyzoite growth, Parasite, 24 (2017) 1–11.
  • [3] Ashwinder K., Kho M.T., Chee P.M., et al., Targeting Heat Shock Proteins 60 and 70 of Toxoplasma gondii as a Potential Drug Target: In Silico Approach, Interdiscip. Sci. Comput. Life Sci., 8 (2015) 374–387.
  • [4] de Barros, R. A. M., Torrecilhas, A. C., Marciano, M. A. M., Mazuz, M. L., Pereira-Chioccola, V. L., & Fux, B. Toxoplasmosis in human and animals around the world. Diagnosis and perspectives in the one health approach. Acta tropica, (2022) 231, 106432.
  • [5] Angel, S. O., Vanagas, L., & Alonso, A. M. (2024). Mechanisms of adaptation and evolution in Toxoplasma gondii, Molecular and Biochemical Parasitology, (2024), 258, 111615.
  • [6] Tutar L., Tutar Y., Heat Shock Proteins; An Overview, Curr. Pharm. Biotechnol., 11 (2010) 216–222.
  • [7] Sherman M.Y., Gabai V.L., Hsp70 in cancer: Back to the future, Oncogene, (2014) 1–9.
  • [8] Tran N.T., Jakovlic I., Wang W.M., In silico characterisation, homology modelling and structure-based functional annotation of blunt snout bream (Megalobrama amblycephala) Hsp70 and Hsc70 proteins, J. Anim. Sci. Technol., 57 (2015) 1–9.
  • [9] Sharma D., Masison D.C., Hsp70 structure, function, regulation and influence on yeast prions, Protein Pept. Lett., 16 (2009) 571–81.
  • [10] Tutar Y., Okan Ş., Heat shock protein 70 purification and characterization from Cyprinion macrastomus and Garra rufa obtusa, J. Therm. Biol., 37 (2012) 95–99.
  • [11] Guruprasad K., Reddy B.V.B., Pandit M.W., Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence, Protein Eng. Des. Sel., 4 (1990) 155–161.
  • [12] Islam M.S., Shahik S.M., Sohel M., et al., In Silico Structural and Functional Annotation of Hypothetical Proteins of Vibrio cholerae O139, Genomics Inform., 13 (2015) 53–59.
  • [13] Evans C.G., Chang L., Gestwick J.E., Heat Shock Protein 70 (Hsp70) as an Emerging Drug Target, J. Med. Chem., 53 (2010) 4585–4602.
  • [14] Tutar L., Is Hsp70 a folder or a modulator protein?, Basic Clin. Sci., (2010) 18–24.
  • [15] Pallavi R., Roy N., Nageshan R.K., et al., Heat Shock Protein 90 as a Drug Target against Protozoan Infections, J. Biol. Chem., 285 (2010) 37964–37975.
  • [16] Mammari N., Halabi, Mohamad Adnan Yaacoub S., Chlala H., Dardé, Marie-Laure Courtioux B., Toxoplasma gondii Modulates the Host Cell Responses: An Overview of Apoptosis Pathways, Biomed. Res. Int., (2019) 1–10.
  • [17] Metzger D.C.H., Hemmer-Hansen J., Schulte P.M., Conserved structure and expression of Hsp70 paralogs in teleost fishes, Comp. Biochem. Physiol. Part D Genomics Proteomics, 18 (2016) 10–20.
  • [18] Luengo T.M., Mayer M.P., Rüdiger S.G.D., The Hsp70 – Hsp90 Chaperone Cascade in Protein Folding, Trends Cell Biol., (2018) 1–14.
  • [19] Mayer M.P., Bukau B., Hsp70 chaperones : Cellular functions and molecular mechanism, Cell Mol. Life Sci., 62 (2005) 670–684.
There are 19 citations in total.

Details

Primary Language English
Subjects Analytical Biochemistry, Protein Trafficking, Medical Biochemistry - Proteins, Peptides and Proteomics
Journal Section Natural Sciences
Authors

Kübra Açıkalın Coşkun 0000-0002-0047-8368

Lütfi Tutar 0000-0002-6260-3136

Publication Date September 30, 2024
Submission Date June 15, 2024
Acceptance Date September 15, 2024
Published in Issue Year 2024

Cite

APA Açıkalın Coşkun, K., & Tutar, L. (2024). Isolation and Characterization of Heat Shock Protein 70-Ipek 1 from Toxoplasma gondii. Cumhuriyet Science Journal, 45(3), 496-502. https://doi.org/10.17776/csj.1501849