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Inhibitory regulation of cystic fibrosis transmembrane conductance regulator anion-transporting activities by Shank2

 Joo Young Kim  ;  WonSun Han  ;  Min Goo Lee  ;  Eunjoon Kim  ;  Hyewon Shin  ;  Kyung Hwan Kim  ;  Ji Hyun Lee  ;  Wan Namkung 
 JOURNAL OF BIOLOGICAL CHEMISTRY, Vol.279(11) : 10389-10396, 2004 
Journal Title
Issue Date
Animals ; Anions* ; Biotinylation ; Cell Membrane/metabolism ; Chlorine/metabolism ; Cyclic AMP/metabolism ; Cystic Fibrosis Transmembrane Conductance Regulator/metabolism ; Cystic Fibrosis Transmembrane Conductance Regulator/physiology* ; Dose-Response Relationship, Drug ; Hydrogen-Ion Concentration ; Immunoblotting ; Immunohistochemistry ; Mice ; Microscopy, Fluorescence ; Mutagenesis, Site-Directed ; NIH 3T3 Cells ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/physiology* ; Oligonucleotides, Antisense/chemistry ; Pancreatic Ducts/metabolism ; Phosphorylation ; Precipitin Tests ; Protein Binding ; Protein Structure, Tertiary ; Protein Transport ; RNA, Messenger/metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Distribution ; Transfection ; Two-Hybrid System Techniques
Accumulating evidence suggests that protein-protein interactions play an important role in transepithelial ion transport. In the present study, we report on the biochemical and functional association between cystic fibrosis transmembrane conductance regulator (CFTR) and a PDZ domain-containing protein Shank2. Exploratory reverse transcription-PCR screening revealed mRNAs for several members of PDZ domain-containing proteins in epithelial cells. Shank2, one of these scaffolding proteins, showed a strong interaction with CFTR by yeast two-hybrid assays. Shank2-CFTR interaction was verified by co-immunoprecipitation experiments in mammalian cells. Notably, this interaction was abolished by mutations in the PDZ domain of Shank2. Protein phosphorylation, HCO(3)(-) transport and Cl(-) current by CFTR were measured in NIH 3T3 cells with heterologous expression of Shank2. Of interest, expression of Shank2 suppressed cAMP-induced phosphorylation and activation of CFTR. Importantly, loss of Shank2 by stable transfection of antisense-hShank2 plasmid strongly increased CFTR currents in colonic T84 cells, in which CFTR and Shank2 were natively expressed. Our results indicate that Shank2 negatively regulates CFTR and that this may play a significant role in maintaining epithelial homeostasis under normal and diseased conditions such as those presented by secretory diarrhea.
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1. College of Medicine (의과대학) > Dept. of Pharmacology (약리학교실) > 1. Journal Papers
2. College of Dentistry (치과대학) > Dept. of Oral Biology (구강생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Kyung Hwan(김경환)
Kim, Joo Young(김주영) ORCID logo https://orcid.org/0000-0003-2623-1491
Lee, Min Goo(이민구) ORCID logo https://orcid.org/0000-0001-7436-012X
Lee, Ji Hyun(이지현)
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