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Ligand Binding and G-protein Coupling of the Serotonin1A Receptor in Cholesterol-enriched Hippocampal Membranes

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Bioscience Reports

Abstract

The serotonin1A receptor is the most extensively studied member of the family of seven transmembrane domain G-protein coupled serotonin receptors. Since a large portion of such transmembrane receptors remains in contact with the membrane lipid environment, lipid–protein interactions assume importance in the structure-function analysis of such receptors. We have earlier reported the requirement of cholesterol for serotonin1A receptor function in native hippocampal membranes by specific depletion of cholesterol using methyl- β-cyclodextrin. In this paper, we monitored the serotonin1A receptor function in membranes that are enriched in cholesterol using a complex prepared from cholesterol and methyl-β-cyclodextrin. Our results indicate that ligand binding and receptor/G-protein interaction of the serotonin1A receptor do not exhibit significant difference in native and cholesterol-enriched hippocampal membranes indicating that further enrichment of cholesterol has little functional consequence on the serotonin1A receptor function. These results therefore provide new information on the effect of cholesterol enrichment on the hippocampal serotonin1A receptor function.

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Abbreviations

BCA:

bicinchoninic acid

DMPC:

dimyristoyl-sn-glycero-3-phosphocholine

GPCR:

G-protein coupled receptor

GTP-γ-S:

guanosine-5′-O-(3-thiotriphosphate)

MβCD:

methyl-β-cyclodextrin

5-HT:

5-hydroxytryptamine (serotonin)

5-HT1A receptor:

5-hydroxytryptamine-1A receptor

8-OH-DPAT:

8-hydroxy-2(di-N-propylamino) tetralin

p-MPPI:

4-(2′-methoxy)-phenyl-1-[2′-(N-2′′-pyridinyl)-p-iodobenzamido] ethyl-piperazine

PMSF:

phenylmethylsulfonyl fluoride

p-MPPF:

4-(2′-methoxy)-phenyl-1-[2′-(N-2′′-pyridinyl)-p-fluorobenzamido]ethyl-piperazine

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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 National Brain Research Centre, Government of India, for the award of a Postdoctoral Fellowship. A.C. is an honorary faculty member of the Jawaharlal Nehru Centre for Advanced Scientific Research, Bangalore (India). We thank S. Rajanna, Shanti Kalipatnapu, Yamuna Devi Paila and Sandeep Shrivastava for help with the tissue collection and members of our laboratory for critically reading the manuscript.

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

    Amitabha Chattopadhyay, Md. Jafurulla & Thomas J. Pucadyil

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  1. Amitabha Chattopadhyay
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  2. Md. Jafurulla
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  3. Thomas J. Pucadyil
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Correspondence to Amitabha Chattopadhyay.

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Chattopadhyay, A., Jafurulla, M. & Pucadyil, T.J. Ligand Binding and G-protein Coupling of the Serotonin1A Receptor in Cholesterol-enriched Hippocampal Membranes. Biosci Rep 26, 79–87 (2006). https://doi.org/10.1007/s10540-006-9009-9

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  • DOI: https://doi.org/10.1007/s10540-006-9009-9

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