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Bioscience Reports

, Volume 16, Issue 4, pp 327–341 | Cite as

Effect of transmembrane Ca2+ gradient on the coupling of β-adrenergic receptors and adenylyl cyclase

  • G. F. Fan
  • X. Y. Yang
  • Y. G. Huang
  • F. Y. Yang
Short Papers
  • 6 Downloads

Abstract

In order to investigate the effect of transmembrane Ca2+ gradient on Gs mediated coupling of β-AR and adenylyl cyclase, β-AR from duck erythrocytes and Gs and adenylyl cyclase from bovine brain cortices were co-reconstituted into asolectin liposomes with different transmembrane Ca2+ gradient. These proteoliposomes were proven to be impermeable to water-soluble substances. The results obtained indicate that a physiological transmembrane Ca2− gradient (1000-fold) is essential for higher stimulation of adenylyl cyclase by hormone-activated β-AR via coupling to Gs and can be further enhanced by the decrease of such Ca2+ gradient within certain range (100 fold) following Ca2+ influx into cells during signal transduction. Fluorescence polarization of DPH revealed that transmembrane Ca2+ gradient modulates adenylyl cyclase and its stimulation by hormones through mediating a change in lipid fluidity. Correspondent conformational changes of β-AR were also detected from the fluorescence spectra and quenching of Acrylodan-labelled β-AR in those proteoliposomes. It is suggested that a proper transmembrane Ca2+ gradient is essential for the optimal fluidity of the phospholipid bilayer in the proteoliposomes, which favors the formation of a suitable conformation of the reconstituted β-AR and thus promotes the stimulation of adenylyl cyclase activities by hormone-activated β-AR via Gs.

Key words

Transmembrane Ca2+ gradient β-adrenergic receptors (β-AR) stimulatory GTP-binding protein (GS) adenylyl cyclase lipid fluidity 

Abbreviations

ATP

adenosine triphosphate

β-AR

β-adrenergic receptors

AC

adenylyl cyclase

DHA

dihydroalprenolol

DPH

diphenylhexatriene

[Ca2+]i

Ca2+ concentration inside proteoliposomes

[Ca2+]o

Ca2+ concentration outside proteoliposomes

cAMP

cyclic adenosine monophosphate

DTT

Dithiothreitol

FS

fluorescein sulfonate

Gs

Stimulatory GTP-binding protein

GTP

guanosine triphosphate

GTPγS

guanosine 5′-O-(3-thiotriphosphate)

kDa

kilodalton

SDS

sodium dodecyl sulfate

Tris

N-tris(hydroxymethyl)aminomethane

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Copyright information

© Plenum Publishing Corporation 1996

Authors and Affiliations

  • G. F. Fan
    • 1
  • X. Y. Yang
    • 1
  • Y. G. Huang
    • 1
  • F. Y. Yang
    • 1
  1. 1.National Laboratory of BiomacromoleculesInstitute of Biophysics, Chinese Academy of SciencesBeijingChina

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