Low levels of expression of leptin receptor at the cell surface result from constitutive endocytosis and intracellular retention in the biosynthetic pathway.
The leptin receptor is mainly localized in intracellular compartments in target tissues. To study the mechanisms leading to this intracellular localization, two main isoforms of leptin receptors, OB-Ra and OB-Rb, were expressed in HeLa cells. Both isoforms were localized at steady state in the trans-Golgi network, in endosomes, and to a lesser extent, at the cell surface. They turned over with a half-life of less than 2 h. Both isoforms of leptin receptors were constitutively endocytosed in a ligand-independent manner and degraded in lysosomes with no evidence of recycling to the cell surface or to the trans-Golgi network. The endocytosis was inhibited by the deletion of the cytoplasmic domain. Newly synthesized leptin receptors were partially retained in the Golgi complex or in a post-Golgi intracellular compartment. The transmembrane domain was found to be important for this intracellular retention in the biosynthetic pathway, whereas the cytoplasmic domain was not involved. The data suggest that the low levels of expression of leptin receptors at the cell surface results from partial retention in the biosynthetic pathway, coupled to constitutive removal from the plasma membrane via ligand-independent, constitutive endocytosis.
Belouzard
Sandrine
S
Unité Propre de Recherche 2511, Institut de Biologie de Lille, CNRS, 59021 Lille, France.
Delcroix
Delphine
D
Rouillé
Yves
Y
D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005822:Genetic Vectors; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D020738:Leptin / Q000737:chemistry; D008024:Ligands; D016296:Mutagenesis; D016297:Mutagenesis, Site-Directed; D011233:Precipitin Tests; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000737:chemistry; D054411:Receptors, Leptin; D013997:Time Factors
C515448:LEPR protein, human; D020738:Leptin; D008024:Ligands; D020033:Protein Isoforms; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin