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

  • Sarojini R. Bulbule
  • P. Aravind
  • N. Hemalatha
  • K. S. Devaraju
Article
  • 16 Downloads

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.

Keywords

Calcineurin (CaN) Protein kinase-A (PKA) Docking Peptide Enzyme assay 

Notes

Acknowledgements

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

Compliance with Ethical Standards

Conflict of interest

Authors contributed to the writing of the manuscript and approved the final version and declare that they have no conflict of interest.

Research Involving Human and Animal Rights

This article does not contain studies with human or animal subjects performed by any of the authors that should be approved by Ethics Committee. The article does not contain any studies in patients by any of the authors.

Informed Consent

The article does not contain any studies in patients by any of the authors.

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Sarojini R. Bulbule
    • 1
  • P. Aravind
    • 1
  • N. Hemalatha
    • 2
  • K. S. Devaraju
    • 1
  1. 1.Department of BiochemistryKarnatak UniversityDharwadIndia
  2. 2.Department of Biochemistry and NutritionCFTRIMysoreIndia

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