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Yeast two-hybrid screening for proteins that interact with alpha(1)-adrenergic receptors
Zhang, T; Xu, Q; Chen, FR; Han, QD; Zhang, YY
关键词Active Bcr-related Protein Alpha(1)-adrenergic Receptors Bone Morphogenetic Protein-1 Filamin-c Yeasts Two-hybrid System Techniques Beta-galactosidase
刊名ACTA PHARMACOLOGICA SINICA
2004-11-01
25期:11页:1471-1478
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Multidisciplinary ; Pharmacology & Pharmacy
研究领域[WOS]Chemistry ; Pharmacology & Pharmacy
关键词[WOS]EHLERS-DANLOS SYNDROME ; GUANINE-NUCLEOTIDE EXCHANGE ; PROCOLLAGEN C-PROTEINASE ; DBL ONCOGENE PRODUCT ; ACTIN CROSS-LINKING ; GAMMA-FILAMIN ; COLLAGEN GENE ; V COLLAGEN ; COL5A1 ; MUSCLE
英文摘要

AIM: To find novel proteins that may bind to alpha(1)-adrenergic receptor (alpha(1B)-AR) and investigate their interactions with the other two alpha(1)-AR subtypes (alpha(1B)-AR and alpha(1D)-AR) with an expectation to provide new leads for the function study of the receptors. METHODS: Yeast two-hybrid assay was performed to screen a human brain cDNA library using the C terminus of alpha(1A)-AR (alpha(1A)-AR-CT) as bait. X-Gal assay and o-nitrophenyl-beta-D-galactopyranoside (ONPG) assay were subsequently conducted to further qualitatively or quantitatively confirm the interactions between receptors and the three identified proteins. RESULTS: (1) Selection medium screening identified segments of bone morphogenetic protein-1 (BMP-1), active Bcr-related protein (Abr), and filamin-C as binding partners of alpha(1A)-AR-CT in yeast cells respectively. Besides, protein segments of BMP-1 and Abr could only specifically interact with alpha(1A)-AR-CT while filamin-C segment interacted with all three alpha(1)-AR subtypes. (2) In X-Gal assay, the cotransformants of alpha(1A)-AR-CT and BMP-1 segments turned strong blue at about 30 min while other positive transformants only developed weak blue at about 5-6 h. (3) In ONPG assay, interaction (shown in beta-galactosidase activity) between alpha(1A)-AR-CT and BMP-1 segments was about 30 times stronger than that of control (P<0.01), while other positive interactions were only about 2-5 times as strong as those of controls (P<0.05). CONCLUSION: In yeast cells BMP-1, Abr and/or filamin-C could interact with three alpha(1)-AR subtypes, among which, interaction between BMP-1 and alpha(1)-AR was the strongest while other interactions between proteins and receptors were relatively weak.

语种英语
WOS记录号WOS:000224947400014
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被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/57184
专题北京大学第三临床医学院_心血管内科
作者单位1.Peking Univ, Hosp 3, Inst Vasc Med, Beijing 100083, Peoples R China
2.Peking Univ, Hosp 3, Key Lab Mol Cardiovasc Sci, Minist Educ, Beijing 100083, Peoples R China
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GB/T 7714
Zhang, T,Xu, Q,Chen, FR,et al. Yeast two-hybrid screening for proteins that interact with alpha(1)-adrenergic receptors[J]. ACTA PHARMACOLOGICA SINICA,2004,25(11):1471-1478.
APA Zhang, T,Xu, Q,Chen, FR,Han, QD,&Zhang, YY.(2004).Yeast two-hybrid screening for proteins that interact with alpha(1)-adrenergic receptors.ACTA PHARMACOLOGICA SINICA,25(11),1471-1478.
MLA Zhang, T,et al."Yeast two-hybrid screening for proteins that interact with alpha(1)-adrenergic receptors".ACTA PHARMACOLOGICA SINICA 25.11(2004):1471-1478.
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