title: RACK1 is a BKCa channel binding protein.
abstract: The large conductance calcium-activated potassium channel, or BK(Ca) channel, plays an important feedback role in a variety of physiological processes, including neurotransmitter release and smooth muscle contraction. Some reports have suggested that this channel forms a stable complex with regulators of its function, including several kinases and phosphatases. To further define such signaling complexes, we used the yeast two-hybrid system to screen a human aorta cDNA library for proteins that bind to the BK(Ca) channel's intracellular, COOH-terminal "tail". One of the interactors we identified is the protein receptor for activated C kinase 1 (RACK1). RACK1 is a member of the WD40 protein family, which also includes the G protein beta-subunits. Consistent with an important role in BK(Ca)-channel regulation, RACK1 has been shown to be a scaffolding protein that interacts with a wide variety of signaling molecules, including cSRC and PKC. We have confirmed the interaction between RACK1 and the BK(Ca) channel biochemically with GST pull-down and coimmunoprecipitation experiments. We have observed some co-localization of RACK1 with the BK(Ca) channel in vascular smooth muscle cells with immunocytochemical experiments, and we have found that RACK1 has effects on the BK(Ca) channel's biophysical properties. Thus RACK1 binds to the BK(Ca) channel and it may form part of a BK(Ca)-channel regulatory complex in vascular smooth muscle.
authors.affiliation: Molecular Cardiology Research Institute, Tufts-New England Medical Center, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
authors.forename: Christina Kaldany
authors.initials: CK
authors.lastname: Isacson
authors.forename: Qing
authors.initials: Q
authors.lastname: Lu
authors.forename: Richard H
authors.initials: RH
authors.lastname: Karas
authors.forename: Daniel H
authors.initials: DH
authors.lastname: Cox
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D005260:Female; D019204:GTP-Binding Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / Q000235:genetics / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000502:physiology*; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology*; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000502:physiology*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D051381:Rats; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D014982:Xenopus laevis
chemical_list: C494841:KCNMA1 protein, human; C494842:Kcnma1 protein, rat; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits; D009363:Neoplasm Proteins; D015221:Potassium Channels; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D019204:GTP-Binding Proteins