title: The kinase domains of obscurin interact with intercellular adhesion proteins.
abstract: Obscurins comprise a family of giant (~870- to 600-kDa) and small (~250- to 55-kDa) proteins that play important roles in myofibrillogenesis, cytoskeletal organization, and cell adhesion and are implicated in hypertrophic cardiomyopathy and tumorigenesis. Giant obscurins are composed of tandem structural and signaling motifs, including 2 serine/threonine kinase domains, SK1 and SK2, present at the COOH terminus of giant obscurin-B. Using biochemical and cellular approaches, we show for the first time that both SK1 and SK2 possess enzymatic activities and undergo autophosphorylation. SK2 can phosphorylate the cytoplasmic domain of N-cadherin, a major component of adherens junctions, and SK1 can interact with the extracellular domain of the β1-subunit of the Na(+)/K(+)-ATPase, which also resides in adherens junctions. Immunostaining of nonpermeabilized myofibers and cardiocytes revealed that some obscurin kinase isoforms localize extracellularly. Quantification of the exofacial expression of obscurin kinase proteins indicated that they occupy ~16 and ~5% of the sarcolemmal surface in myofibers and cardiocytes, respectively. Treatment of heart lysates with peptide-N-glycosidase F revealed that while giant obscurin-B localizes intracellularly, possessing dual kinase activity, a small obscurin kinase isoform that contains SK1 localizes extracellularly, where it undergoes N-glycosylation. Collectively, our studies demonstrate that the obscurin kinase domains are enzymatically active and may be involved in the regulation of cell adhesion.
authors: Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
authors: Li-Yen R
authors: LY
authors: Hu
authors: Aikaterini
authors: A
authors: Kontrogianni-Konstantopoulos
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D005260:Female; D006031:Glycosylation; D020662:Guanine Nucleotide Exchange Factors / Q000276:immunology / Q000378:metabolism*; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D009124:Muscle Proteins / Q000276:immunology / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000201:enzymology; D009210:Myofibrils / Q000201:enzymology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000502:physiology; D064067:Rho Guanine Nucleotide Exchange Factors; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: C503552:Atp1b1 protein, mouse; D015820:Cadherins; C489083:Cdh2 protein, mouse; D020662:Guanine Nucleotide Exchange Factors; D007527:Isoenzymes; D009124:Muscle Proteins; D064067:Rho Guanine Nucleotide Exchange Factors; D017346:Protein Serine-Threonine Kinases; C515766:obscn protein, mouse; D000254:Sodium-Potassium-Exchanging ATPase
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