title: The cAMP-dependent protein kinase phosphorylates the rap1 protein in vitro as well as in intact fibroblasts, but not the closely related rap2 protein.
abstract: The products of rap genes (rap1A, rap1B and rap2) are small molecular weight GTP-binding proteins that share approximately 50% homology with ras-p21s. It had previously been shown that a rap1 protein (also named Krev-1 or smg p21) could be phosphorylated on serine residues by the cAMP-dependent protein kinase (PKA) in vitro as well as in intact platelets stimulated by prostaglandin E1. We show here that the rap1A protein purified from recombinant bacteria is phosphorylated in vitro by the catalytic subunit of PKA and that the deletion of the 17 C-terminal amino acids leads to the loss of this phosphorylation. This suggests that the serine residue at position 180 constitutes the site of phosphorylation of the rap1A protein by PKA. The rap1 protein can also be phosphorylated by PKA in intact fibroblasts; this phenomenon is independent of their proliferative state. In contrast, protein kinase C (PKC) does not phosphorylate the rap1 proteins, neither in vitro nor in vivo. Finally, the 60% homologous rap2 protein is neither phosphorylated in vitro nor in vivo by PKA or PKC.
authors.affiliation: INSERM U-248, Faculté de Médecine Lariboisière-Saint Louis, Paris, France.
authors.forename: I
authors.initials: I
authors.lastname: Lerosey
authors.forename: V
authors.initials: V
authors.lastname: Pizon
authors.forename: A
authors.initials: A
authors.lastname: Tavitian
authors.forename: J
authors.initials: J
authors.lastname: de Gunzburg
mesh_terms: D000818:Animals; D002455:Cell Division; D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D011493:Protein Kinase C / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D020668:rap GTP-Binding Proteins
chemical_list: D011994:Recombinant Proteins; D011494:Protein Kinases; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D020668:rap GTP-Binding Proteins