La α2-macroglobulina del Ovis aries, su purificación y la comparación con las α2-macroglobulinas humanas
Resumen
Purificar la α2-Macroglobulina del plasma del Ovis aries y compararla con las α-macroglobulinas y en particular con la α2-Macroglobulina humana. Materiales y métodos: la α2-Macroglobulina humana fue purificada por cromatografía de afinidad y la α2-Macroglobulina del Ovis aries, por precipitaciones repetidas de polietilenglicol 6000, seguido de diálisis y cromatografías de cambio iónico y exclusión. Ambas proteínas fueron caracterizadas por electroforesis SDS-PAGE y nativa, dichas proteínas fueron incubadas con quimotripsina en las proporciones 1:1, 1:2, 1:4 respectivamente, las secuencias determinadas por el método de Edman. Resultados: La purificación de las proteínas alcanzó una pureza mayor al 95% determinada por densitometría, en la electroforesis denaturante se demostró una sola banda de 180 kDa, para cada una de las proteínas y en el gel de electroforesis nativa, se visualizó una banda en cada proteína que migraron paralelas. En la determinación de las secuencias para las α-Macroglobulinas humanas y de Ovis aries, se encontraron varias similitudes entre ellas Tyr-Met-Val-Leu-Val, otras secuencias determinadas en las α-Macroglobulinas mostraron menor similitud. Electroforéticamente se demostró, las mismas características en ambas proteínas (humano y Ovis aries). Se demostró la forma rápida y lenta en la α2-Macroglobulina de Ovis aries con quimotripsina, típicas formas en la reacción de la α2-Macroglobulina humana con proteinasas. Se confirmó por determinación de secuencia, que esta proteína pertenece al grupo de las α-Macroglobulinas.
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DOI: https://doi.org/10.24054/01204211.v2.n2.2013.843
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