Cloning and expression of the Ca2+ channel α(1C) and β(2a) subunits from guinea pig heart

Shan Ding, Sachiko Kuroki, Asako Kameyama, Akihiko Yoshimura, Masaki Kameyama

研究成果: Article

11 引用 (Scopus)

抄録

Complimentary DNA clones encoding the α(1C) and β(2a) subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2169 amino acids for the α(1C) and 597 amino acids for the β(2a) subunit. The proteins showed 94.2 and 94.8%, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig α(1C) and β(2a) subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the α(1C) subunit is expressed exclusively in the heart, while the β(2a) subunit is expressed in the heart, cerebellum, whole brain, and stomach. The α(1C) and β(2a) subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing α(1C) alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of β(2a) with α(1C) did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig α(1C) and rabbit β1 + α2/δ, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced S-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.

元の言語English
ページ(範囲)750-759
ページ数10
ジャーナルJournal of Biochemistry
125
発行部数4
出版物ステータスPublished - 1999
外部発表Yes

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Cloning
Organism Cloning
Guinea Pigs
3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester
Amino Acids
Okadaic Acid
Clamping devices
Colforsin
Nifedipine
Brain
Proteins
Cells
Modulation
DNA
Rabbits
Electric potential
Polymerase Chain Reaction
Protein Subunits
Patch-Clamp Techniques
Cardiac Myocytes

ASJC Scopus subject areas

  • Biochemistry

これを引用

Cloning and expression of the Ca2+ channel α(1C) and β(2a) subunits from guinea pig heart. / Ding, Shan; Kuroki, Sachiko; Kameyama, Asako; Yoshimura, Akihiko; Kameyama, Masaki.

:: Journal of Biochemistry, 巻 125, 番号 4, 1999, p. 750-759.

研究成果: Article

Ding, S, Kuroki, S, Kameyama, A, Yoshimura, A & Kameyama, M 1999, 'Cloning and expression of the Ca2+ channel α(1C) and β(2a) subunits from guinea pig heart', Journal of Biochemistry, 巻. 125, 番号 4, pp. 750-759.
Ding, Shan ; Kuroki, Sachiko ; Kameyama, Asako ; Yoshimura, Akihiko ; Kameyama, Masaki. / Cloning and expression of the Ca2+ channel α(1C) and β(2a) subunits from guinea pig heart. :: Journal of Biochemistry. 1999 ; 巻 125, 番号 4. pp. 750-759.
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abstract = "Complimentary DNA clones encoding the α(1C) and β(2a) subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2169 amino acids for the α(1C) and 597 amino acids for the β(2a) subunit. The proteins showed 94.2 and 94.8{\%}, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig α(1C) and β(2a) subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the α(1C) subunit is expressed exclusively in the heart, while the β(2a) subunit is expressed in the heart, cerebellum, whole brain, and stomach. The α(1C) and β(2a) subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing α(1C) alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of β(2a) with α(1C) did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig α(1C) and rabbit β1 + α2/δ, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced S-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.",
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T1 - Cloning and expression of the Ca2+ channel α(1C) and β(2a) subunits from guinea pig heart

AU - Ding, Shan

AU - Kuroki, Sachiko

AU - Kameyama, Asako

AU - Yoshimura, Akihiko

AU - Kameyama, Masaki

PY - 1999

Y1 - 1999

N2 - Complimentary DNA clones encoding the α(1C) and β(2a) subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2169 amino acids for the α(1C) and 597 amino acids for the β(2a) subunit. The proteins showed 94.2 and 94.8%, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig α(1C) and β(2a) subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the α(1C) subunit is expressed exclusively in the heart, while the β(2a) subunit is expressed in the heart, cerebellum, whole brain, and stomach. The α(1C) and β(2a) subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing α(1C) alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of β(2a) with α(1C) did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig α(1C) and rabbit β1 + α2/δ, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced S-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.

AB - Complimentary DNA clones encoding the α(1C) and β(2a) subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2169 amino acids for the α(1C) and 597 amino acids for the β(2a) subunit. The proteins showed 94.2 and 94.8%, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig α(1C) and β(2a) subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the α(1C) subunit is expressed exclusively in the heart, while the β(2a) subunit is expressed in the heart, cerebellum, whole brain, and stomach. The α(1C) and β(2a) subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing α(1C) alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of β(2a) with α(1C) did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig α(1C) and rabbit β1 + α2/δ, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced S-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.

KW - α(1C) sununit

KW - β subunit

KW - Ca channel

KW - Cloning

KW - Guinea pig heart

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M3 - Article

C2 - 10101289

AN - SCOPUS:0033113113

VL - 125

SP - 750

EP - 759

JO - Journal of Biochemistry

JF - Journal of Biochemistry

SN - 0021-924X

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