Design and validation of a homogeneous time-resolved fluorescence-based leptin receptor binding assay (original) (raw)

“…ObR are mainly retained in the Golgi, trans-Golgi network and endosomes, whereas the surface receptor undergoes permanent constitutive endocytosis, independent of leptin stimulation, leading to lysosomal degradation without apparent recycling [57]. These processes result in short receptor half-life (less than 2 h) and in low levels of both ObRa and ObRb isoforms at the cell surface (5–20%) [57, 58]. The level of 5–20% of ObR at the plasma membrane does not constitute the maximal activation capacity of leptin signaling.…”

Section: Central Leptin Resistance Associated With Obesitymentioning

“…[41,49,50]). The K D is mostly in the range of 10 -3 -10 -9 M. One example for a fluorescent ligand binding validation study that concerns the GMP-regulated quality assessment has been published by de Boer et al [51].…”

Section: Fluorescence Based Ligand Binding Assaysmentioning

“…At early times during gestation when the BBB is not yet fully functional, blood-borne factors may be able to gain access via free diffusion or some other process that is not present in the mature BBB. Tagging of peripheral factors, as has been done with leptin in the adult brain (Vauthier et al , 2013; Balland, 2014), may be one approach to resolving this important question (for example, see Banks et al , 2004). …”

Section: Prenatal Dietary Programming Of Behavior and Immunity Acrossmentioning