Serine and Threonine Phosphorylation of the Low Density Lipoprotein Receptor-related Protein by Protein Kinase Cα Regulates Endocytosis and Association with Adaptor Molecules

• Published in: The Journal of biological chemistry Vol. 279; no. 39; p. 40536
• Main Authors: Sripriya Ranganathan, Chun-Xiang Liu, Mary M. Migliorini, Christine A. F. von Arnim, Ithan D. Peltan, Irina Mikhailenko, Bradley T. Hyman, Dudley K. Strickland
• Format: Journal Article
• Language: English
• Published: American Society for Biochemistry and Molecular Biology 24.09.2004
• DOI: 10.1074/jbc.M407592200
• ISSN: 0021-9258

The low density lipoprotein receptor-related protein (LRP) is a large receptor that participates in endocytosis, signaling pathways, and phagocytosis of necrotic cells. Mechanisms that direct LRP to function in these distinct pathways likely involve its association with distinct cytoplasmic adaptor proteins. We tested the hypothesis that the association of various adaptor proteins with the LRP cytoplasmic domain is modulated by its phosphorylation state. Phosphoamino acid analysis of metabolically labeled LRP revealed that this receptor is phosphorylated at serine, threonine, and tyrosine residues within its cytoplasmic domain, whereas inhibitor studies identified protein kinase Cα (PKCα) as a kinase capable of phosphorylating LRP. Mutational analysis identified critical threonine and serine residues within the LRP cytoplasmic domain that are necessary for phosphorylation mediated by PKCα. Mutating these threonine and serine residues to alanines generated a receptor that was not phosphorylated and that was internalized more rapidly than wild-type LRP, revealing that phosphorylation reduces the association of LRP with adaptor molecules of the endocytic machinery. In contrast, serine and threonine phosphorylation was necessary for the interaction of LRP with Shc, an adaptor protein that participates in signaling events. Furthermore, serine and threonine phosphorylation increased the interaction of LRP with other adaptor proteins such as Dab-1 and CED-6/GULP. These results indicate that phosphorylation of LRP by PKCα modulates the endocytic and signaling function of LRP by modifying its association with adaptor proteins.