@article{APS8409,
author = {Tan ZHANG and Qi XU and Fen-rong CHEN and Qi-de HAN and You-yi ZHANG},
title = {Yeast two-hybrid screening for proteins that interact with alpha1-adrenergic receptors},
journal = {Acta Pharmacologica Sinica},
volume = {25},
number = {11},
year = {2016},
keywords = {},
abstract = {AIM:
To find novel proteins that may bind to alpha1A-adrenergic receptor (alpha1A-AR) and investigate their interactions with the other two alpha1-AR subtypes (alpha1B-AR and alpha1D-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 alpha1A-AR (alpha1A-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 alpha1A-AR-CT in yeast cells respectively. Besides, protein segments of BMP-1 and Abr could only specifically interact with alpha1A-AR-CT while filamin-C segment interacted with all three alpha1-AR subtypes. (2) In X-Gal assay, the co-transformants of alpha1A-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 alpha1A-AR-CT and BMP-1 segments was about 30 times stronger than that of control (P},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/8409}
}