RRC ID 72583
Author Takano M, Kanoh A, Amako K, Otani M, Sano K, Kanazawa-Hamada M, Matsuyama S.
Title Nuclear localization of bradykinin B(2) receptors reflects binding to the nuclear envelope protein lamin C.
Journal Eur J Pharmacol
Abstract The mechanism of action of bradykinin (BK), a pro-inflammatory mediator, is thought to be mediated by specific cell surface membrane bradykinin B2 receptors. Some evidence suggests that there are both intracellular and nuclear bradykinin B2 receptors. This study identified proteins that interact with the C-terminus of the bradykinin B2 receptor (in particular, the nuclear membrane protein lamin C), using the yeast two-hybrid system. The motif of the C-terminal domain (CT) mutant 303-320 in bradykinin B2 receptor was identified as a lamin C protein binding motif. Immunohistochemistry revealed colocalization of FLAG- bradykinin B2 receptor with HA-lamin C in the nucleus of HEK 293T cells. In situ proximity ligation assay (PLA) showed that FLAG-bradykinin B2 receptor formed heterodimers with HA-lamin C in the nucleus. In addition, live cell fluorescence imaging showed that bradykinin B2 receptor-EGFP was located in the nucleus and co-localized with HcRed-lamin C. Interestingly, neither BK addition nor bradykinin B2 receptor CT mutation reduced the binding to lamin C or changed the distribution of bradykinin B2 receptor. Taken together, these findings demonstrate that bradykinin B2 receptor-lamin C heterodimers form in the nucleus independent of BK stimulation and CT mutation. We propose that heterodimerization of bradykinin B2 receptor with lamin C is essential to nuclear localization of bradykinin B2 receptor and plays an important role in cell signaling and function.
Volume 723
Pages 507-14
Published 2014-1-15
DOI 10.1016/j.ejphar.2013.09.054
PII S0014-2999(13)00718-8
PMID 24211782
MeSH Animals Cell Nucleus / metabolism DNA, Complementary / genetics HEK293 Cells Humans Lamin Type A / genetics Lamin Type A / metabolism* Mice Mutation Receptor, Bradykinin B2 / genetics Receptor, Bradykinin B2 / metabolism* Two-Hybrid System Techniques
IF 3.263
Human and Animal Cells 293T(RCB2202)