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Molecular Pharmacology

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Research ArticleArticle

Mammalian Tolloid Alters Subcellular Localization, Internalization, and Signaling of α1a-Adrenergic Receptors

Qi Xu, Ning Xu, Tan Zhang, Hui Zhang, Zijian Li, Feng Yin, Zhizhen Lu, Qide Han and Youyi Zhang
Molecular Pharmacology August 2006, 70 (2) 532-541; DOI: https://doi.org/10.1124/mol.105.016451
Qi Xu
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Ning Xu
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Tan Zhang
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Hui Zhang
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Zijian Li
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Feng Yin
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Zhizhen Lu
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Qide Han
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Youyi Zhang
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Abstract

In the present study, we identified the CUB5 domain of mammalian Tolloid (mTLD) as a novel protein binding to α1A-adrenergic receptor (AR) using the yeast two-hybrid system. Whereas CUB5 did not couple to either α1B-AR or α1D-AR. It was determined that amino acids 322 to 359 of α1A-AR were the major binding region for CUB5. The direct interaction between α1A-AR cytoplasmic tail and CUB5 was discovered by glutathione S-transferase pull-down assay. We confirmed the interaction of mTLD with α1A-AR in human embryonic kidney (HEK) 293 cells by immunoprecipitation, immunofluorescence, and fluorescence resonance energy transfer. Although mTLD did not affect the density and affinity of receptors in crudely prepared membranes from HEK293 cells stably expressing α1A-AR, it significantly altered the subcellular localization of the receptors. Moreover, mTLD reduced the level of cell surface α1A-ARs, delayed the initial rate of agonist-induced receptor internalization, and facilitated agonist-induced calcium transient. We have demonstrated that mTLD interacts with α1A-AR directly, alters the subcellular localization of receptor, and influences agonist-induced α1A-AR internalization and calcium signaling.

  • Received July 4, 2005.
  • Accepted May 11, 2006.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 70 (2)
Molecular Pharmacology
Vol. 70, Issue 2
1 Aug 2006
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Research ArticleArticle

Mammalian Tolloid Alters Subcellular Localization, Internalization, and Signaling of α1a-Adrenergic Receptors

Qi Xu, Ning Xu, Tan Zhang, Hui Zhang, Zijian Li, Feng Yin, Zhizhen Lu, Qide Han and Youyi Zhang
Molecular Pharmacology August 1, 2006, 70 (2) 532-541; DOI: https://doi.org/10.1124/mol.105.016451

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Research ArticleArticle

Mammalian Tolloid Alters Subcellular Localization, Internalization, and Signaling of α1a-Adrenergic Receptors

Qi Xu, Ning Xu, Tan Zhang, Hui Zhang, Zijian Li, Feng Yin, Zhizhen Lu, Qide Han and Youyi Zhang
Molecular Pharmacology August 1, 2006, 70 (2) 532-541; DOI: https://doi.org/10.1124/mol.105.016451
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