Similar ligand binding in recombinant human α2C2-adrenoceptors produced in mammalian, insect and yeast cells

Anne Marjamäki; Katariina Pohjanoksa; Sari Ala-Uotila; Dorothea Sizmann; Christian Oker-Blom; Hitoshi Kurose; Mika Scheinin

doi:10.1016/0922-4106(94)90232-1

Similar ligand binding in recombinant human α₂C2-adrenoceptors produced in mammalian, insect and yeast cells

Anne Marjamäki
, Katariina Pohjanoksa
, Sari Ala-Uotila
, Dorothea Sizmann
, Christian Oker-Blom
, Hitoshi Kurose
, Mika Scheinin

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

Ligand binding properties were investigated in recombinant human α₂C2-adrenoceptors expressed in three different host systems: Shionogi S115 mouse mammary tumour cells, Spodoptera frugiperda Sf9 insect cells and Saccharomyces cerevisae yeast cells. The expected 43 kDa α₂C2 protein was visualized with immunoblotting using a polyclonal α₂C2-receptor antibody. [³H]Rauwolscine binding in cell homogenates or membranes (B_max 3-11 pmol/mg protein; K_d approximately 5.5 nM) was inhibited by prazosin, oxymetazoline, RX821002, chlorpromazine and (-)-noradrenaline with and without the GTP-analogue Gpp(NH)p with similar K_i values in the different host systems. This indicates that α₂C2-adrenoceptors retain their binding characteristics irrespective of the host environment.

Original language	English
Pages (from-to)	117-121
Number of pages	5
Journal	European Journal of Pharmacology: Molecular Pharmacology
Volume	267
Issue number	1
DOIs	https://doi.org/10.1016/0922-4106(94)90232-1
Publication status	Published - Mar 15 1994
Externally published	Yes

All Science Journal Classification (ASJC) codes

Pharmacology

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10.1016/0922-4106(94)90232-1

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title = "Similar ligand binding in recombinant human α2C2-adrenoceptors produced in mammalian, insect and yeast cells",

abstract = "Ligand binding properties were investigated in recombinant human α2C2-adrenoceptors expressed in three different host systems: Shionogi S115 mouse mammary tumour cells, Spodoptera frugiperda Sf9 insect cells and Saccharomyces cerevisae yeast cells. The expected 43 kDa α2C2 protein was visualized with immunoblotting using a polyclonal α2C2-receptor antibody. [3H]Rauwolscine binding in cell homogenates or membranes (Bmax 3-11 pmol/mg protein; Kd approximately 5.5 nM) was inhibited by prazosin, oxymetazoline, RX821002, chlorpromazine and (-)-noradrenaline with and without the GTP-analogue Gpp(NH)p with similar Ki values in the different host systems. This indicates that α2C2-adrenoceptors retain their binding characteristics irrespective of the host environment.",

author = "Anne Marjam{\"a}ki and Katariina Pohjanoksa and Sari Ala-Uotila and Dorothea Sizmann and Christian Oker-Blom and Hitoshi Kurose and Mika Scheinin",

year = "1994",

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doi = "10.1016/0922-4106(94)90232-1",

language = "English",

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journal = "European Journal of Pharmacology: Molecular Pharmacology",

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T1 - Similar ligand binding in recombinant human α2C2-adrenoceptors produced in mammalian, insect and yeast cells

AU - Marjamäki, Anne

AU - Pohjanoksa, Katariina

AU - Ala-Uotila, Sari

AU - Sizmann, Dorothea

AU - Oker-Blom, Christian

AU - Kurose, Hitoshi

AU - Scheinin, Mika

PY - 1994/3/15

Y1 - 1994/3/15

N2 - Ligand binding properties were investigated in recombinant human α2C2-adrenoceptors expressed in three different host systems: Shionogi S115 mouse mammary tumour cells, Spodoptera frugiperda Sf9 insect cells and Saccharomyces cerevisae yeast cells. The expected 43 kDa α2C2 protein was visualized with immunoblotting using a polyclonal α2C2-receptor antibody. [3H]Rauwolscine binding in cell homogenates or membranes (Bmax 3-11 pmol/mg protein; Kd approximately 5.5 nM) was inhibited by prazosin, oxymetazoline, RX821002, chlorpromazine and (-)-noradrenaline with and without the GTP-analogue Gpp(NH)p with similar Ki values in the different host systems. This indicates that α2C2-adrenoceptors retain their binding characteristics irrespective of the host environment.

AB - Ligand binding properties were investigated in recombinant human α2C2-adrenoceptors expressed in three different host systems: Shionogi S115 mouse mammary tumour cells, Spodoptera frugiperda Sf9 insect cells and Saccharomyces cerevisae yeast cells. The expected 43 kDa α2C2 protein was visualized with immunoblotting using a polyclonal α2C2-receptor antibody. [3H]Rauwolscine binding in cell homogenates or membranes (Bmax 3-11 pmol/mg protein; Kd approximately 5.5 nM) was inhibited by prazosin, oxymetazoline, RX821002, chlorpromazine and (-)-noradrenaline with and without the GTP-analogue Gpp(NH)p with similar Ki values in the different host systems. This indicates that α2C2-adrenoceptors retain their binding characteristics irrespective of the host environment.

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U2 - 10.1016/0922-4106(94)90232-1

DO - 10.1016/0922-4106(94)90232-1

M3 - Article

C2 - 7911432

AN - SCOPUS:0028296702

SN - 0922-4106

VL - 267

SP - 117

EP - 121

JO - European Journal of Pharmacology: Molecular Pharmacology

JF - European Journal of Pharmacology: Molecular Pharmacology

IS - 1

ER -

Similar ligand binding in recombinant human α₂C2-adrenoceptors produced in mammalian, insect and yeast cells

Abstract

All Science Journal Classification (ASJC) codes

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