Document Detail


Cloning of a novel protein interacting with BRS-3 and its effects in wound repair of bronchial epithelial cells.
MedLine Citation:
PMID:  21857995     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bombesin receptor subtype 3 (BRS-3), the orphan bombesin receptor, may play a role in the regulation of stress responses in lung and airway epithelia. Bombesin receptor activated protein (BRAP )is a novel protein we found in our previous study which interacts with BRS-3. This study was designed to observe the subcellular location and wound repair function of BRAP in human bronchial epithelial cells (HBECs). BRAP ORF was amplified by RT-PCR and ligated to pEGFP-C1 vector, and then the recombinant plasmid pEGFP-C1-BRAP was transfected into Hela cells. The location of BRAP protein was observed by laser confocal microscope, and the expression of it was analyzed by Western-blot. At the same time,we built the recombinant plasmid pcDNA3.1(+)-BRAP, transfected it into HBECs and observed its impact on cell cycle and wound repair of HBECs. The results showed that BRAP locates in membrane and cytoplasm and increases significantly in transfected cells. Flow cytometry results demonstrated that the recombinant plasmid increases S phase plus G2 phase of cell cycle by 25%. Microscopic video analysis system showed that the repair index of wounded HBECs increases by 20% through stable expression of BRAP. The present study demonstrated that BRAP locates in the membrane and cytoplasm, suggesting that this protein is a cytoplasm protein, which promotes cell cycle and wound repair of HBECs.
Authors:
Hui Jun Liu; Yu Rong Tan; Meng Lan Li; Chi Liu; Yang Xiang; Xiao Qun Qin
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-08-09
Journal Detail:
Title:  PloS one     Volume:  6     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2011  
Date Detail:
Created Date:  2011-08-22     Completed Date:  2012-02-15     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e23072     Citation Subset:  IM    
Affiliation:
Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, China.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Base Sequence
Blotting, Western
Bronchi / cytology,  metabolism
Cell Line
Cell Membrane / metabolism
Cloning, Molecular
Cytoplasm / metabolism
Epithelial Cells / metabolism*
Flow Cytometry
G2 Phase
Green Fluorescent Proteins / genetics,  metabolism
HeLa Cells
Humans
Microscopy, Confocal
Molecular Sequence Data
Protein Binding
Receptors, Bombesin / genetics,  metabolism*
Recombinant Fusion Proteins / genetics,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
S Phase
Transfection
Ubiquitin-Protein Ligases / genetics,  metabolism*
Wound Healing
Chemical
Reg. No./Substance:
0/Receptors, Bombesin; 0/Recombinant Fusion Proteins; 0/bombesin receptor subtype 3; 147336-22-9/Green Fluorescent Proteins; EC 6.3.2.-/BRAP protein, human; EC 6.3.2.19/Ubiquitin-Protein Ligases
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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