Details

Autor(en) / Beteiligte

• Kuwahara, Michio
• Fushimi, Kiyohide
• Terada, Yoshio
• Bai, Liqun
• Marumo, Fumiaki
• Sasaki, Sei

Titel

cAMP-dependent Phosphorylation Stimulates Water Permeability of Aquaporin-collecting Duct Water Channel Protein Expressed in Xenopus Oocytes

Ist Teil von

• The Journal of biological chemistry, 1995-05, Vol.270 (18), p.10384-10387

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

1995

Quelle

MEDLINE

Beschreibungen/Notizen

• Among water channel proteins (aquaporins), aquaporin-collecting duct (AQP-CD) is the vasopressin-regulated water channel. Vasopressin causes cAMP production in the renal collecting duct cells, and this is believed to lead to exocytic insertion of water channel into the apical membrane (shuttle hypothesis). AQP-CD contains a consensus sequence for cAMP-dependent protein kinase, residues at positions 253-256 (Arg- Arg-Gln- Ser). To determine the role of this site, Ser-256 was substituted for Ala, Leu, Thr, Asp, or Glu by site-directed mutagenesis. In Xenopus oocytes injected with wild-type or mutated AQP-CD cRNAs, osmotic water permeability (Pf) was 4.8-7.7 times higher than Pf of water-injected oocytes. Incubation with cAMP plus forskolin or direct cAMP injection into the oocytes increased Pf of wild-type, but not mutated, AQP-CD-expressing oocytes, whereas the amounts of AQP-CD expression were similar in wild and mutated types as identified by Western blot analysis. In vitro phosphorylation studies of AQP-CD proteins expressed in oocyte showed that cAMP-dependent protein kinase phosphorylated wild-type, but not mutated, AQP-CD proteins. Phosphoamino acid analysis revealed that this phosphorylation occurred at the serine residue. Moreover, phosphorylation of AQP-CD protein in intact rat kidney medulla tissues was stimulated by incubation with cAMP. Our data suggest that cAMP stimulates water permeability of AQP-CD by phosphorylation. This process may contribute to the vasopressin-regulated water permeability of collecting duct in addition to the apical insertion of AQP-CD by exocytosis.