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Modeling, Synthesis and Characterization of Phospho–Penta–Peptide derived from PKA RII Sub Unit: A Candidate Substrate for CaN Assay

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Abstract

Calcineurin (CaN) plays an important role in many pathological conditions like cancer, metabolic aberrations and neurodegenerative disorders. Till date contemporary methods to assay CaN uses p-nitrophenylphosphate (p-NPP) and specific protein phosphatases 2B inhibitor or use of phospho-peptide substrate derived from protein kinase A (PKA) regulatory sub unit type PKARII. Here we have modeled a series of small phospho peptides derived from PKARII and were docked by Molegro Virtual Docking software to find out the binding free enegy and to determine the affinity with modeled Calcineurin catalytic subunit A (CaN A). Further the candidate phospho–penta–peptide (PPP) derived from PKA regulatory sub unit type II was synthesized in house by employing Fmoc chemistry by solution phase method, purified over HPLC and characterized by mass spectrometry. This PPP was employed to assay CaN activity by Malachite Green method found to be a candidate substrate for CaN.

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Acknowledgements

Authors acknowledge the financial support DST-SERB, Government of India (Grant No. SB/EMEQ-238/2013).

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Authors and Affiliations

  1. Department of Biochemistry, Karnatak University, Pavate Nagar, Dharwad, 580 003, India

    Sarojini R. Bulbule, P. Aravind & K. S. Devaraju

  2. Department of Biochemistry and Nutrition, CFTRI, V V Moholla, Mysore, 570 020, India

    N. Hemalatha

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  1. Sarojini R. Bulbule
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  2. P. Aravind
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  3. N. Hemalatha
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  4. K. S. Devaraju
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Correspondence to K. S. Devaraju.

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Bulbule, S.R., Aravind, P., Hemalatha, N. et al. Modeling, Synthesis and Characterization of Phospho–Penta–Peptide derived from PKA RII Sub Unit: A Candidate Substrate for CaN Assay. Int J Pept Res Ther 25, 1251–1258 (2019). https://doi.org/10.1007/s10989-018-9770-1

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  • DOI: https://doi.org/10.1007/s10989-018-9770-1

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