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学科主题: 临床医学
题名:
Yeast two-hybrid screening for proteins that interact with alpha(1)-adrenergic receptors
作者: 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
发表日期: 2004-11-01
卷: 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
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/57184
Appears in Collections:北京大学第三临床医学院_心血管内科_期刊论文

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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

Recommended Citation:
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.
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