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The human serotonin1A receptor exhibits G-protein-dependent cell surface dynamics

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Abstract

Seven transmembrane domain G-protein-coupled receptors constitute the largest family of proteins in mammals. Signal transduction events mediated by such receptors are the primary means by which cells communicate with and respond to their external environment. The major paradigm in this signal transduction process is that stimulation of the receptor leads to the recruitment and activation of heterotrimeric GTP-binding proteins. These initial events, which are fundamental to all types of G-protein-coupled receptor signaling, occur at the plasma membrane via protein–protein interactions. As a result, the dynamics of the activated receptor on cell surfaces represents an important determinant in its encounter with G-proteins, and has significant impact on the overall efficiency of the signal transduction process. We have monitored the cell surface dynamics of the serotonin1A receptor, an important member of the G-protein-coupled receptor superfamily, in relation to its interaction with G-proteins. Fluorescence recovery after photobleaching experiments carried out with the receptor tagged to the enhanced yellow fluorescent protein indicate that G-protein activation alters the diffusion properties of the receptor in a manner suggesting the activation process leads to dissociation of G-proteins from the receptor. This result demonstrates that the cell surface dynamics of the serotonin1A receptor is modulated in a G-protein-dependent manner. Importantly, this result could provide the basis for a sensitive and powerful approach to assess receptor/G-protein interaction in an intact cellular environment.

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Abbreviations

5-HT1A receptor:

5-hydroxytryptamine-1A receptor

EYFP:

enhanced yellow fluorescent protein

FRAP:

fluorescence recovery after photobleaching

GFP:

green fluorescent protein

GPCR:

G-protein-coupled receptor

PCR:

polymerase chain reaction

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Acknowledgments

This work was supported by the Council of Scientific and Industrial Research, Government of India. T.J.P. thanks the Council of Scientific and Industrial Research for the award of a Senior Research Fellowship. A.C. is an honorary faculty member of the Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore (India). We thank Dr. Sadashiva S. Karnik (The Cleveland Clinic Foundation, Cleveland, USA) for the construct coding for the 5-HT1A receptor tagged to EYFP, Dr. G. Krishnamoorthy (Tata Institute for Fundamental Research, Mumbai, India) for the kind gift of purified GFP and Dr. Preeti G. Joshi (National Institute of Mental Health and Neurosciences, Bangalore, India) for the kind gift of pertussis toxin. We gratefully acknowledge Nandini Rangaraj, V.K. Sarma, N.R. Chakravarthi, and K.N. Rao for technical assistance in confocal microscopy.

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  1. Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad, 500 007, India

    Thomas J. Pucadyil & Amitabha Chattopadhyay

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  1. Thomas J. Pucadyil
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Correspondence to Amitabha Chattopadhyay.

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Pucadyil, T.J., Chattopadhyay, A. The human serotonin1A receptor exhibits G-protein-dependent cell surface dynamics. Glycoconj J 24, 25–31 (2007). https://doi.org/10.1007/s10719-006-9008-x

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