Structure, Function, and Dynamics of the G&#945; Binding Domain of Ric-8A.

Zeng, Baisen; Mou, Tung-Chung; Doukov, Tzanko I; Steiner, Andrea; Yu, Wenxi; Papasergi-Scott, Makaia; Tall, Gregory G; Hagn, Franz; Sprang, Stephen R

Structure, Function, and Dynamics of the Gα Binding Domain of Ric-8A.

Zeng, Baisen; Mou, Tung-Chung; Doukov, Tzanko I; Steiner, Andrea; Yu, Wenxi; Papasergi-Scott, Makaia; Tall, Gregory G; Hagn, Franz; Sprang, Stephen R.

Afiliación

Zeng B; Graduate Program in Biochemistry and Biophysics, University of Montana, Missoula, MT 59812, USA.
Mou TC; Center for Biomolecular Structure and Dynamics, University of Montana, Missoula, MT 59812, USA; Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA.
Doukov TI; Macromolecular Crystallography Group, Stanford Synchrotron Radiation Light Source, SLAC National Accelerator Laboratory, Stanford University, Stanford, CA 94309, USA.
Steiner A; Bavarian NMR Center at the Department of Chemistry and Institute for Advanced Study, Technical University of Munich, Ernst-Otto-Fischer-Strasse 2, 85748 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
Yu W; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Papasergi-Scott M; Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Tall GG; Department of Pharmacology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
Hagn F; Bavarian NMR Center at the Department of Chemistry and Institute for Advanced Study, Technical University of Munich, Ernst-Otto-Fischer-Strasse 2, 85748 Garching, Germany; Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
Sprang SR; Graduate Program in Biochemistry and Biophysics, University of Montana, Missoula, MT 59812, USA; Center for Biomolecular Structure and Dynamics, University of Montana, Missoula, MT 59812, USA; Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA. Electronic address: stephe

Structure ; 27(7): 1137-1147.e5, 2019 07 02.

Article en En | MEDLINE | ID: mdl-31155309

RESUMEN

Ric-8A is a 530-amino acid cytoplasmic molecular chaperone and guanine nucleotide exchange factor (GEF) for i, q, and 12/13 classes of heterortrimeric G protein alpha subunits (Gα). We report the 2.2-Å crystal structure of the Ric-8A Gα-binding domain with GEF activity, residues 1-452, and is phosphorylated at Ser435 and Thr440. Residues 1-429 adopt a superhelical fold comprised of Armadillo (ARM) and HEAT repeats, and the C terminus is disordered. One of the phosphorylated residues potentially binds to a basic cluster in an ARM motif. Amino acid sequence conservation and published hydrogen-deuterium exchange data indicate repeats 3 through 6 to be a putative Gα-binding surface. Normal mode modeling of small-angle X-ray scattering data indicates that phosphorylation induces relative rotation between repeats 1-4, 5-6, and 7-9. 2D 1H-15N-TROSY spectra of [2H,15N]-labeled Gαi1 in the presence of R452 reveals chemical shift perturbations of the C terminus and Gαi1 residues involved in nucleotide binding.

Asunto(s)

Subunidades alfa de la Proteína de Unión al GTP Gi-Go/química; Factores de Intercambio de Guanina Nucleótido/química; Proteínas Nucleares/química; Proteínas Recombinantes de Fusión/química; Secuencia de Aminoácidos; Animales; Sitios de Unión; Clonación Molecular; Cristalografía por Rayos X; Escherichia coli/genética; Escherichia coli/metabolismo; Subunidades alfa de la Proteína de Unión al GTP Gi-Go/genética; Subunidades alfa de la Proteína de Unión al GTP Gi-Go/metabolismo; Expresión Génica; Vectores Genéticos/química; Vectores Genéticos/metabolismo; Factores de Intercambio de Guanina Nucleótido/genética; Factores de Intercambio de Guanina Nucleótido/metabolismo; Guanosina Trifosfato; Histidina/genética; Histidina/metabolismo; Modelos Moleculares; Mutación; Proteínas Nucleares/genética; Proteínas Nucleares/metabolismo; Oligopéptidos/genética; Oligopéptidos/metabolismo; Unión Proteica; Conformación Proteica en Hélice alfa; Conformación Proteica en Lámina beta; Dominios y Motivos de Interacción de Proteínas; Multimerización de Proteína; Ratas; Proteínas Recombinantes de Fusión/genética; Proteínas Recombinantes de Fusión/metabolismo

Palabras clave

X-ray crystallography; guanine nucleotide exchange factor; heteronuclear nuclear magnetic resonance; heterotrimeric G protein; molecular chaperone; protein dynamics; protein structure; small-angle X-ray scattering

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Recombinantes de Fusión / Proteínas Nucleares / Subunidades alfa de la Proteína de Unión al GTP Gi-Go / Factores de Intercambio de Guanina Nucleótido Tipo de estudio: Prognostic_studies Idioma: En Revista: Structure Asunto de la revista: BIOLOGIA MOLECULAR / BIOQUIMICA / BIOTECNOLOGIA Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

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