Ca(2+)/calmodulin-dependent protein kinase mediates the phosphorylation of CD44 required for cell migration on hyaluronan.
CD44 is the principal cell surface receptor for the extracellular matrix glycosaminoglycan hyaluronan, and binding to this ligand underlies CD44-mediated cell attachment and migration. As would be expected for a widely expressed adhesion receptor, CD44 is subject to complex regulatory events, and mis-regulation of the receptor has been associated with a number of disease pathologies, including chronic inflammatory conditions and the progression of metastatic tumours. In previous studies we have demonstrated that a key control point for this receptor is the phosphorylation of CD44 on a conserved cytoplasmic serine residue, Ser(325). This modification is not required for efficient ligand binding, but is an essential component of CD44-dependent cell migration on a hyaluronan substratum. To understand better the mechanism regulating CD44 phosphorylation on Ser(325), we have generated a monoclonal antibody that specifically recognizes CD44 phosphorylated on Ser(325), and have developed assays to identify the Ser(325) kinase. We demonstrate here that CD44 is phosphorylated to high stoichiometry in resting cells and that Ca(2+)/calmodulin-dependent protein kinase II is a CD44 Ser(325) kinase.
Lewis
C A
CA
Department of Biology, Sir Alexander Fleming Building, Imperial College of Science, Technology and Medicine, Imperial College Road, London SW7 2AZ, UK.
Townsend
P A
PA
Isacke
C M
CM
D000595:Amino Acid Sequence; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism* / Q000502:physiology; D006820:Hyaluronic Acid / Q000502:physiology*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D051381:Rats; D012694:Serine / Q000378:metabolism; D013379:Substrate Specificity
D018960:Hyaluronan Receptors; D012694:Serine; D006820:Hyaluronic Acid; D017871:Calcium-Calmodulin-Dependent Protein Kinases