ES2372709T3 - Procedimientos de cribado de proteasas y proteasas identificadas por los mismos. - Google Patents

Procedimientos de cribado de proteasas y proteasas identificadas por los mismos. Download PDF

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ES2372709T3
ES2372709T3 ES07861330T ES07861330T ES2372709T3 ES 2372709 T3 ES2372709 T3 ES 2372709T3 ES 07861330 T ES07861330 T ES 07861330T ES 07861330 T ES07861330 T ES 07861330T ES 2372709 T3 ES2372709 T3 ES 2372709T3
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proteases
modified
procedures
identified
same processes
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Edwin L. Madison
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Torrey Pines Institute For Molecular Studies
Gyre Therapeutics Inc
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Torrey Pines Institute For Molecular Studies
Catalyst Biosciences Inc
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Abstract

Un procedimiento para identificar o seleccionar una proteasa mutante o porción catalíticamente activa de la misma que escinde una secuencia de escisión en un sustrato diana de proteína que comprende: a) poner en contacto un conjunto de una pluralidad de proteasas mutantes y/o porciones catalíticamente activas de las proteasas mutantes con un polipéptido de captura de proteasas, en el que: el polipéptido de captura de proteasas comprende un sitio reactivo que contiene una secuencia de escisión para el sustrato diana de proteína; el polipéptido de captura de proteasas se selecciona de entre una serpina, un miembro de la familia de las alfa-macroglobulinas, un miembro de la familia p35 y una forma modificada de las mismas que se modifica en el sitio reactivo por uno o más reemplazos, deleciones o sustituciones de aminoácidos para incluir la secuencia de escisión del sustrato diana de proteína; la puesta en contacto se efectúa en condiciones para la escisión de aminoácidos en la secuencia de escisión incluida en el sitio reactivo en el polipéptido de captura de proteasas por una proteasa o una porción catalíticamente activa de la misma en el conjunto para formar complejos estables que contienen el polipéptido de captura de proteasas ligado covalentemente a una proteasa o porción catalíticamente activa de la misma; y tras la escisión de aminoácidos en la secuencia de escisión incluida en el sitio reactivo en el polipéptido de captura de proteasas por una proteasa o porción catalíticamente activa de la misma en el conjunto, el polipéptido de captura de proteasas forma un enlace covalente con la proteasa o porción catalíticamente activa de la misma para formar un complejo estable que contiene una proteasa o porción catalíticamente activa de la misma con un polipéptido de captura de proteasas; b) separar las proteasas complejadas de los miembros sin complejar del conjunto; y c) identificar o seleccionar una proteasa o porción catalíticamente activa de la misma en un complejo, para identificar o seleccionar una proteasa o porción catalíticamente activa de la misma que escinde la secuencia de escisión en el polipéptido de captura de proteasas, identificándose o seleccionándose así una proteasa o porción catalíticamente activa de la misma que escinde el sustrato diana de proteína.
ES07861330T 2006-07-05 2007-07-05 Procedimientos de cribado de proteasas y proteasas identificadas por los mismos. Active ES2372709T3 (es)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US81891006P 2006-07-05 2006-07-05
US81880406P 2006-07-05 2006-07-05
US818910P 2006-07-05
US818804P 2006-07-05

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