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The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain.
González Bardeci, Nicolás; Caramelo, Julio J; Blumenthal, Donald K; Rinaldi, Jimena; Rossi, Silvia; Moreno, Silvia.
Afiliação
  • González Bardeci N; Departamento de Química Biológica, IQUIBICEN-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina.
  • Caramelo JJ; Departamento de Química Biológica, IQUIBICEN-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina; Fundación Instituto Leloir, IIBBA-CONICET, C1405BWE Buenos Aires, Argentina.
  • Blumenthal DK; Department of Pharmacology & Toxicology, University of Utah, Salt Lake City, UT 84112, USA.
  • Rinaldi J; Fundación Instituto Leloir, IIBBA-CONICET, C1405BWE Buenos Aires, Argentina.
  • Rossi S; Departamento de Química Biológica, IQUIBICEN-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina.
  • Moreno S; Departamento de Química Biológica, IQUIBICEN-CONICET, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, C1428EGA Buenos Aires, Argentina. Electronic address: smoreno@qb.fcen.uba.ar.
J Struct Biol ; 193(2): 141-54, 2016 Feb.
Article em En | MEDLINE | ID: mdl-26687415
The cAMP dependent protein kinase (PKA) is a key enzyme involved in many cellular processes in eukaryotes. In mammals, the regulatory (R) subunit localises the catalytic (C) subunit to specific subcellular sites through the interaction of its N-terminal homodimeric docking and dimerization (D/D) domain with specific scaffold proteins. The structure of the D/D domain has been extensively studied in mammals, but there is little information from non-mammalian species. In this work, we present the structural analysis of the D/D domain of Bcy1, the R subunit of PKA from Saccharomyces cerevisiae. Using chemical crosslinking experiments and static light scattering measurements we found that this R subunit forms a tetramer in solution, unlike its dimeric mammalian counterparts. We determined that the D/D domain is responsible for this unusual oligomeric state. Using biophysical techniques including size-exclusion chromatography, sucrose gradient sedimentation, small angle X-ray scattering (SAXS), and circular dichroism, we performed a detailed structural characterization of the tetrameric D/D domain of Bcy1. We used homology modelling in combination with computer-aided docking methods and ab initio SAXS modelling methods to develop structural models for the D/D domain tetramer. The models consist of two homodimers with a canonical D/D domain fold that generate a dimer of dimers with novel putative interaction surfaces. These findings indicate that the oligomerization states of PKA R subunits is more diverse than previously thought, and suggest that this might allow some forms of PKA to interact with a wide range of intracellular partners.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Quinases Dependentes de AMP Cíclico / Proteínas de Saccharomyces cerevisiae Tipo de estudo: Prognostic_studies Idioma: En Revista: J Struct Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos
Texto completo
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XML
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Quinases Dependentes de AMP Cíclico / Proteínas de Saccharomyces cerevisiae Tipo de estudo: Prognostic_studies Idioma: En Revista: J Struct Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos