Skip to main content
Research article - Peer-reviewed, 2006

Purification and characterization of two protein kinases acting on the aquaporin SoPIP2; 1

Sjövall-Larsen, Sara; Alexandersson, Erik; Johansson, Ingela; Karlsson, Maria; Johanson, Urban; Kjellbom, Per


Aquaporins are water channel proteins that facilitate the movement of water and other small solutes across biological membranes. Plants usually have large aquaporin families, providing them with many ways to regulate the water transport. Some aquaporins are regulated post-translationally by phosphorylation. We have previously shown that the water channel activity of SoP1P2;1, an aquaporin in the plasma membrane of spinach leaves, was enhanced by phosphorylation at Ser115 and Ser274. These two serine residues are highly conserved in all plasma membrane aquaporins of the PIP2 subgroup. In this study we have purified and characterized two protein kinases phosphorylating Ser115 and Ser274 in SoPIP2;1. By anion exchange chromatography, the Ser115 kinase was purified from the soluble protein fraction isolated from spinach leaves. The Ca2+-dependent Ser274 kinase was purified by peptide affinity chromatography using plasma membranes isolated from spinach leaves. When characterized, the Ser115 kinase was Mg2+-dependent, Ca2+-independent and had a pH-optimum at 6.5. In accordance with previous studies using the oocyte expression system, site-directed mutagenesis and kinase and phosphatase inhibitors, the phosphorylation of Ser274, but not of Ser115, was increased in the presence of phosphatase inhibitors while kinase inhibitors decreased the phosphorylation of both Ser274 and Ser115. The molecular weight of the Ser274 kinase was approximately 50 kDa. The identification and characterization of these two protein kinases is an important step towards elucidating the signal transduction pathway for gating of the aquaporin SoPIP2;1. (c) 2006 Elsevier B.V. All rights reserved.


aquaporm; PIP2; SoPIP2; 1; phosphorylation; protein kinase

Published in

BBA - Biomembranes
2006, Volume: 1758, number: 8, pages: 1157-1164

    SLU Authors

    UKÄ Subject classification

    Biochemistry and Molecular Biology

    Publication Identifiers


    Permanent link to this page (URI)