Translocation of telokin by cGMP signaling in smooth muscle cells
Department of Physiology
Animals; Binding Sites; COS Cells; Cell Membrane; Cells, Cultured; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Enzyme Inhibitors; Green Fluorescent Proteins; Luminescent Proteins; Muscle Proteins; Muscle, Smooth; Mutagenesis, Site-Directed; Myosin-Light-Chain Kinase; Myosins; Nitric Oxide Donors; Peptides; Phosphorylation; Protein Binding; Protein Transport; Recombinant Fusion Proteins; Signal Transduction; Swine; Trachea; Transfection
Life Sciences | Medicine and Health Sciences
Telokin is an acidic protein with a sequence identical to the COOH-terminal domain of myosin light chain kinase (MLCK) produced by an alternate promoter of the MLCK gene. Although it is abundantly expressed in smooth muscle, its physiological function is not understood. In the present study, we attempted to clarify the function of telokin by analyzing its spatial and temporal localization in living single smooth muscle cells. Primary cultured smooth muscle cells were transfected with green fluorescent protein (GFP)-tagged telokin. The telokin-GFP localized mostly diffusely in cytosol. Stimulation with both sodium nitroprusside (SNP) and 8-bromo-cyclic GMP induced translocation of GFP-tagged telokin to near plasma membrane in living single smooth muscle cells. The translocation was slow, and it took more than 10 min at room temperature. Mutation of the phosphorylation sites of telokin (S13A, S19A, and S13A/S19A) significantly attenuated SNP-induced translocation. Both KT-5823 (cGMP-dependent protein kinase inhibitor) and PD-98059 (mitogen-activated protein kinase inhibitor) diminished the telokin-GFP translocation. These results suggest that telokin changes its intracellular localization because of phosphorylation at Ser13 and/or Ser19 via the cGMP-signaling pathway.
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Citation: Am J Physiol Cell Physiol. 2002 Sep;283(3):C752-61. Link to article on publisher's site
DOI of Published Version
American journal of physiology. Cell physiology
Komatsu, Satoshi; Miyazaki, Koji; Tuft, Richard A.; and Ikebe, Mitsuo, "Translocation of telokin by cGMP signaling in smooth muscle cells" (2002). Open Access Articles. 115.