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Metabolomics pp 161–178Cite as

Assays for Intracellular Cyclic Adenosine Monophosphate (cAMP) and Lysosomal Acidification

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1996))

Abstract

Cyclic adenosine monophosphate (3′,5′-cAMP) is a multifunctional second messenger which controls extremely diverse and physiologically important biochemical pathways. Among its myriad roles, 3′,5′-cAMP functions as an intracellular regulator of lysosomal pH, which is essential for the activity of acidic lysosomal enzymes. Defects in lysosomal acidification are attributed to many diseases like macular degeneration, Parkinson’s, Alzheimer’s, and cystic fibrosis. Strategic re-acidification of defective lysosomes by pharmacological increase of intracellular cAMP offers exciting therapeutic potential in these diseases. Modular assays for accurate assessment of intracellular cAMP and lysosomal pH are a critical component of this research. We describe label-free targeted metabolomics for quantitating intracellular cAMP and integrated assays for measuring lysosomal pH. These hybrid assays offer fast, unbiased information on intracellular cAMP concentrations and lysosomal pH that can be applied to many cell types and putative drug screening strategies.

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Acknowledgments

The authors gratefully acknowledge Mr. Dipankar Malakar, Sciex(India), for assistance with the data analysis. The program was supported by the Centre for Advanced Research grant from the Indian Council of Medical Research and DBT-Twinning Research Grant, from the Department of Biotechnology, Government of India, to DG.

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Authors and Affiliations

  1. Special Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, India

    Chiranjit Maity & Dipankar Ghosh

  2. Stein Eye Institute, University of California Los Angeles, Los Angeles, CA, USA

    Sonia Guha

Authors
  1. Chiranjit Maity
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  2. Dipankar Ghosh
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  3. Sonia Guha
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Correspondence to Sonia Guha .

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Editors and Affiliations

  1. Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA

    Sanjoy K. Bhattacharya

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Maity, C., Ghosh, D., Guha, S. (2019). Assays for Intracellular Cyclic Adenosine Monophosphate (cAMP) and Lysosomal Acidification. In: Bhattacharya, S. (eds) Metabolomics. Methods in Molecular Biology, vol 1996. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9488-5_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9488-5_14

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