The Modular Adaptor Protein ARH Is Required for Low Density Lipoprotein (LDL) Binding and Internalization but Not for LDL Receptor Clustering in Coated Pits

• Published in: The Journal of biological chemistry Vol. 279; no. 32; pp. 34023 - 34031
• Main Authors: Michaely, Peter, Li, Wei-Ping, Anderson, Richard G.W., Cohen, Jonathan C., Hobbs, Helen H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 06.08.2004; American Society for Biochemistry and Molecular Biology
• Subjects: Adaptor Proteins, Signal Transducing; Adaptor Proteins, Vesicular Transport - deficiency; Adaptor Proteins, Vesicular Transport - physiology; Cell Line, Transformed; Cell Membrane - chemistry; Coated Pits, Cell-Membrane - chemistry; Coated Pits, Cell-Membrane - metabolism; Endocytosis; Fluorescent Antibody Technique; Humans; Hypercholesterolemia - blood; Immunohistochemistry; Iodine Radioisotopes; Lipoproteins, LDL - blood; Lymphocytes - ultrastructure; Microscopy, Electron; Receptors, LDL - analysis; Receptors, LDL - blood
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M405242200

ARH is an adaptor protein required for efficient endocytosis of low density lipoprotein (LDL) receptors (LDLRs) in selected tissues. Individuals lacking ARH (ARH-/-) have severe hypercholesterolemia due to impaired hepatic clearance of LDL. Immortalized lymphocytes, but not fibroblasts, from ARH-deficient subjects fail to internalize LDL. To further define the role of ARH in LDLR function, we compared the subcellular distribution of the LDLR in lymphocytes from normal and ARH-/- subjects. In normal lymphocytes LDLRs were predominantly located in intracellular compartments, whereas in ARH-/- cells the receptors were almost exclusively on the plasma membrane. Biochemical assays and quantification of LDLR by electron microscopy indicated that ARH-/- lymphocytes had >20-fold more LDLR on the cell surface and a ∼27-fold excess of LDLR outside of coated pits. The accumulation of LDLR on the cell surface was not due to failure of receptors to localize in coated pits since the number of LDLRs in coated pits was similar in ARH-/- and normal cells. Despite the dramatic increase in cell surface receptors, LDL binding was only 2-fold higher in the ARH-/- lymphocytes. These findings indicate that ARH is required not only for internalization of the LDL·LDLR complex but also for efficient binding of LDL to the receptor and suggest that ARH stabilizes the associations of the receptor with LDL and with the invaginating portion of the budding pit, thereby increasing the efficiency of LDL internalization.