The C2 cytosolic loop of adenylyl cyclase interacts with the activated form of Gαs

FEBS Letters

Genevière, Anne-Marie¹  Echeverrı́a, Valentina²  Guzmán, Leonardo²  Olate, Juan¹  Hinrichs, Marı́a Victoria²  Torrejón, Marcela² 

[1] Observatoire Océanologique de Banyuls, Laboratoire Arago, Université Pierre et Marie Curie (Paris VI), Institut National des Sciences de L'Univers, Banyuls-Sur-Mer Cedex, France;Laboratorio de Genética Molecular, Departamento de Biologı́a Molecular, Facultad de Ciencias Biológicas, Casilla 160-C, Universidad de Concepción, Concepción, Chile

关键词: Adenylyl cyclase;    G protein;    Yeast two-hybrid system;   

DOI  :  10.1016/S0014-5793(98)01603-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using the yeast two-hybrid system, we studied the physical interaction between the complete C₁ and C₂ cytosolic domains of Xenopus laevis type 9 (xl9C₁, xl9C₂) and the C₂ domain of rat type 6 (r6C₂) adenylyl cyclase (AC). Heterodimerization between xl9C₁ and xl9C₂ and homodimerization between C₂ (but not C₁) domains was observed. Interaction between C₂ and human Gαs (hGαs) was also detected and was dependent on Gαs activation. In contrast X. laevis Gαs (xlGαs), which is 92% identical to hGαs, was unable to interact with any of the three AC cytosolic domains tested, corroborating previous findings that showed no effector activation. Through the construction of chimeras, we demonstrated that the amino-terminal half of xlGαs was responsible for the lack of interaction with AC. Chimeras between mouse Gαi2 and Gαs (N-mGαi2/C-Gαs), that have previously shown to activate AC to a higher extent than wild-type Gαs, also interacted with the C₂ cytosolic domain and with a higher affinity. Interestingly, N-mGαi2/C-xlGαs chimera was not only able to interact with C₂ but also with the C₁ cytosolic domain.

【 授权许可】

Unknown