The hinge region of the scaffolding protein of cell contacts, zonula occludens protein 1, regulates interacting with various signaling proteins

被引:8
作者
Bal, Manjot Singh [1 ]
Castro, Victor [1 ]
Piontek, Joerg [1 ]
Rueckert, Christine [1 ]
Walter, Juliane K. [1 ]
Shymanets, Aliaksei [3 ,4 ]
Kurig, Barbara [3 ,4 ]
Haase, Hannelore [2 ]
Nuernberg, Bernd [3 ,4 ]
Blasig, Ingolf E. [1 ]
机构
[1] Leibniz Inst Mol Pharmacol, D-13125 Berlin, Germany
[2] Max Delbruck Ctr Mol Med, Berlin, Germany
[3] Univ Tubingen, Interfac Ctr Pharmacogen & Drug Res ICePhA, Tubingen, Germany
[4] Univ Tubingen, Dept Pharmacol & Expt Therapy, Inst Expt & Clin Pharmacol & Toxicol, Tubingen, Germany
关键词
CELL-CELL CONTACTS; PROTEIN-PROTEIN INTERACTION; REGULATORY PROTEINS; SURFACE PLASMON RESONANCE SPECTROSCOPY; PEPTIDE BINDING ASSAYS; G-PROTEINS; TIGHT-JUNCTION; CALCIUM-CHANNEL; ZO-1; KINASE; AHNAK; INVOLVEMENT; RECEPTOR; BINDING; PHOSPHORYLATES; DIMERIZATION;
D O I
10.1002/jcb.23422
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Zonula occludens protein 1 (ZO-1) is a ubiquitous scaffolding protein, but it is unknown why it functions in very different cellular contacts. We hypothesized that a specific segment, the unique hinge region, can be bound by very different regulatory proteins. Using surface plasmon resonance spectroscopy and binding assays to peptide libraries, we show, for the first time, that the hinge region directly interacts with disparate signal elements such as G-proteins alpha 12 and alpha i2, the regulator of G-protein signaling 5, multifunctional signaling protein ahnak1, and L-type Ca2+-channel beta-2-subunit. The novel binding proteins specifically bound to a coiled coil-helix predicted in the hinge region of ZO-1. The interactions were modulated by phosphorylation in the hinge helix. Activation of the G-proteins influenced their association to ZO-1. In colon cells, G alpha i2 and ZO-1 were associated, as shown by coimmunoprecipitation. After cotransfection in kidney cells, G alpha i2 barely colocalized with ZO-1; the colocalization coefficient was significantly increased when epinephrine activated G-protein signaling. In conclusion, proteins with different regulatory potential adhere to and influence cellular functions of ZO-proteins, and the interactions can be modulated via its hinge region and/or the binding proteins. J. Cell. Biochem. 113: 934945, 2012. (C) 2011 Wiley Periodicals, Inc.
引用
收藏
页码:934 / 945
页数:12
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