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La clonación de la expresión de un receptor de calcitonina acoplado a adenilato ciclasa acoplado a la adenilato
H Y Lin1, T L Harris, M S Flannery
1Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Science (New York, N.Y.)
|November 25, 1991
Resumen
Los investigadores clonaron un ADN complementario del receptor de la calcitonina (ADNc). Este nuevo receptor, funcionalmente acoplado a la señalización cíclica de adenosina monofosfato (cAMP), representa una nueva familia de receptores acoplados a proteínas G involucrados en la homeostasis del calcio.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Endocrinología Endocrinología.
- La bioquímica es la bioquímica.
Sus antecedentes:
- La calcitonina es una hormona clave que regula la homeostasis del calcio.
- La identidad molecular y la familia del receptor de la calcitonina eran previamente desconocidas.
- La comprensión de los receptores hormonales reguladores del calcio es crucial para la investigación metabólica.
Objetivo del estudio:
- Clonar y caracterizar el ADN complementario (ADNc) del receptor de la calcitonina.
- Para determinar el acoplamiento funcional y las vías de señalización del receptor de la calcitonina.
- Para investigar la relación evolutiva del receptor de la calcitonina con otros receptores acoplados a proteínas G.
Principales métodos:
- Clonado del cDNA del receptor de la calcitonina utilizando la expresión en células COS de una línea celular epitelial de riñón de cerdo.
- Ensayos vinculantes para determinar la afinidad por la calcitonina del salmón (Kd).
- Medición de los niveles intracelulares de adenosina monofosfato cíclico (cAMP) para evaluar el acoplamiento funcional.
Principales resultados:
- Se clonó con éxito un cDNA receptor de calcitonina de 482 aminoácidos.
- El receptor exhibió una alta afinidad por la calcitonina del salmón (Kd ≈ 6 nM).
- Se demostró un acoplamiento funcional al aumento de los niveles intracelulares de cAMP.
Conclusiones:
- El receptor de calcitonina clonado representa un nuevo receptor acoplado a la proteína G.
- Muestra homología con el receptor de péptido relacionado con la hormona paratiroidea y la hormona paratiroidea (PTH-PTHrP), lo que sugiere una nueva familia de receptores.
- Este descubrimiento proporciona información sobre los mecanismos moleculares de la regulación de la homeostasis del calcio.
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