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La clonación molecular y la expresión de un receptor para el factor de necrosis tumoral humano

T J Schall¹, M Lewis, K J Koller

¹Department of Molecular Biology, Genentech, Inc., South San Francisco, California 94080.

|April 20, 1990

Resumen

Los investigadores identificaron una proteína de unión soluble al factor de necrosis tumoral (TNF) en el suero de pacientes con cáncer. Esta proteína probablemente se deriva del receptor TNF (TNF-R), una molécula de la superficie celular crucial para la señalización del TNF.

Área de la Ciencia:

Inmunología Inmunología.
Biología Molecular Biología Molecular
La bioquímica es la bioquímica.

Sus antecedentes:

El factor de necrosis tumoral (TNF) es una citoquina clave involucrada en la inflamación y la inmunidad.
Se han detectado proteínas solubles que se unen al TNF en el suero de pacientes con cáncer, pero su origen no estaba claro.

Objetivo del estudio:

Para purificar y caracterizar una proteína humana de unión al TNF del suero.
Identificar el gen que codifica esta proteína de unión y dilucidar su relación con el receptor TNF.

Principales métodos:

Purificación de proteínas y secuenciación parcial.
Mutagenesis dirigida por oligonucleótidos y clonación de ADNc.
Pruebas de transfección celular de mamíferos y ensayos de unión al receptor.
Análisis de la mancha del norte.

Principales resultados:

Se purificó una proteína homogénea de unión al TNF del suero de pacientes con cáncer.
Se aislaron clones de ADNc que codifican un receptor TNF (TNF-R).
El TNF-R es una proteína transmembrana de 415 aminoácidos con homología al receptor NGF.
Las células transfectadas que expresan TNF-R se unen específicamente al TNF-alfa, y esta unión fue inhibida por el TNF-alfa y el TNF-beta no etiquetados.
El análisis del norte reveló una sola especie de ARNm TNF-R en varios tipos de células.

Conclusiones:

La proteína soluble de unión al TNF que se encuentra en el suero humano es probablemente un fragmento liberado proteolíticamente del receptor del TNF.
Este hallazgo proporciona una base molecular para la presencia de proteínas solubles de unión al TNF en pacientes con cáncer y tiene implicaciones para la señalización mediada por el TNF.

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