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Study of adenylyl cyclase-GαS interactions and identification of novel AC ligands

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Abstract

Adenylyl cyclases (ACs) are membrane bound enzymes that catalyze the production of cAMP from ATP in response to the activation by G-protein Gαs. Different isoforms of ACs are ubiquitously expressed in different tissues involved in regulatory mechanisms in response to specific stimulants. There are 9 AC isoforms present in humans, with AC5 and AC6 proposed to play a vital role in cardiac functions. The activity of AC6 is sensitive to nitric oxide, such that nitrosylation of the protein might regulate its function. However, the information on structural determinants of nitrosylation in ACs and how they interact with Gαs is limited. Here we used homology modeling to build a molecular model of human AC6 bound to Gαs. Based on this 3D model, we predict the nitrosylation amenable cysteines, and identify potential novel ligands of AC6 using virtual ligand screening. Our model suggests Cys1004 in AC6 (subunit C2) and Cys174 in Gαs present at the AC-Gαs interface as the possible residues that might undergo reversible nitrosylation. Docking analysis predicted novel ligands of AC6 that include forskolin-based compounds and its derivatives. Further work involving site-directed mutagenesis of the predicted residues will allow manipulation of AC activity using novel ligands, and crucial insights on the role of nitrosylation of these proteins in pathophysiological conditions.

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Acknowledgements

The work was supported by operating grants from Natural Sciences and Engineering Research Council (NSERC) to PC, Research Manitoba (RM) and Manitoba Chemosensory Biology (MCSB) catalyst grant to SD and University of Manitoba (Faculty of Science Interdisciplinary Grant) to JLS.

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Authors and Affiliations

  1. Manitoba Chemosensory Biology (MCSB) Research Group, Winnipeg, MB, R3E 0W4, Canada

    Appalaraju Jaggupilli, Premnath Dhanaraj, Alexander Pritchard, John L. Sorensen, Shyamala Dakshinamurti & Prashen Chelikani

  2. Department of Oral Biology, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Appalaraju Jaggupilli, Premnath Dhanaraj & Prashen Chelikani

  3. Department of Chemistry, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Alexander Pritchard & John L. Sorensen

  4. Departments of Pediatrics, Physiology, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Shyamala Dakshinamurti

  5. Biology of Breathing, Children’s Hospital Research Institute of Manitoba (CHRIM), Winnipeg, MB, R3E 3P4, Canada

    Shyamala Dakshinamurti & Prashen Chelikani

  6. Department of Oral Biology, and Manitoba Chemosensory Biology (MCSB) Research Group, D319, University of Manitoba, Winnipeg, MB, R3E 0W4, Canada

    Prashen Chelikani

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  1. Appalaraju Jaggupilli
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Correspondence to Prashen Chelikani.

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Jaggupilli, A., Dhanaraj, P., Pritchard, A. et al. Study of adenylyl cyclase-GαS interactions and identification of novel AC ligands. Mol Cell Biochem 446, 63–72 (2018). https://doi.org/10.1007/s11010-018-3273-4

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  • DOI: https://doi.org/10.1007/s11010-018-3273-4

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