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Identification of residues in the N terminus of alpha 1B critical for inhibition of the voltage-dependent calcium channel by G beta gamma

Canti, C; Page, KM; Stephens, GJ; Dolphin, AC; (1999) Identification of residues in the N terminus of alpha 1B critical for inhibition of the voltage-dependent calcium channel by G beta gamma. J NEUROSCI , 19 (16) 6855 - 6864.

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Abstract

To examine the role of the intracellular N terminus in the G-protein modulation of the neuronal voltage-dependent calcium channel (VDCC) alpha 1B, we have pursued two routes of investigation. First, we made chimeric channels between alpha 1B and alpha 1C, the latter not being modulated by G beta gamma subunits. VDCC alpha 1 subunit constructs were coexpressed with accessory alpha 2 delta and beta 2a subunits in Xenopus oocytes and mammalian (COS-7) cells. G-protein modulation of expressed alpha 1 subunits was induced by activation of coexpressed dopamine (D2) receptors with quinpirole in oocytes, or by cotransfection of G beta 1 gamma 2 subunits in COS-7 cells. For the chimeric channels, only those with the N terminus of alpha 1B showed any G-protein modulation; further addition of the first transmembrane domain and I-II intracellular linker of alpha 1B increased the degree of modulation. To determine the amino acids within the alpha 1B N terminus, essential for G-protein modulation, we made mutations of this sequence and identified three amino acids (S48, R52, and R54) within an 11 amino acid sequence as being critical for G-protein modulation, with I49 being involved to a lesser extent. This sequence may comprise an essential part of a complex G beta gamma-binding site or be involved in its subsequent action.

Type: Article
Title: Identification of residues in the N terminus of alpha 1B critical for inhibition of the voltage-dependent calcium channel by G beta gamma
Keywords: calcium channel, neuronal, G-protein, alpha 1 subunit, G beta gamma subunit, modulation, G-PROTEIN MODULATION, CA2+ CHANNELS, DOMAIN-I, INTRACELLULAR LOOP, ALPHA(1) SUBUNIT, BINDING, FACILITATION, CELLS, DETERMINANTS, EXPRESSION
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology
URI: http://discovery.ucl.ac.uk/id/eprint/131320
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