Design and validation of a homogeneous time-resolved fluorescence-based leptin receptor binding assay

Vauthier, Virginie; Derviaux, Carine; Douayry, Najim; Roux, Thomas; Trinquet, Eric; Jockers, Ralf; Dam, Julie

doi:10.1016/j.ab.2012.12.013

Cited by 20 publications

(17 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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

confidence: 99%

Endospanin 1 Determines the Balance of Leptin-Regulated Hypothalamic Functions

2018

Self Cite

View full text Add to dashboard Cite

Endospanin 1 (Endo1), a protein encoded in humans by the same gene than the leptin receptor (ObR), and increased by diet-induced obesity, is an important regulator of ObR trafficking and cell surface exposure, determining leptin signaling strength. Defective intracellular trafficking of the leptin receptor to the neuronal plasma membrane has been proposed as a mechanism underlying the development of leptin resistance observed in human obesity. More recently, Endo1 has emerged as a mediator of “selective leptin resistance.” The underlying mechanisms of the latter are not completely understood, but the possibility of differential activation of leptin signaling pathways was suggested among others. In this respect, the expression level of Endo1 is crucial for the appropriate balance between different leptin signaling pathways and leptin functions in the hypothalamus and is likely participating in selective leptin resistance for the control of energy and glucose homeostasis.

show abstract

Section: Central Leptin Resistance Associated With Obesitymentioning

confidence: 99%

Endospanin 1 Determines the Balance of Leptin-Regulated Hypothalamic Functions

2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…[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

confidence: 99%

Data quality in drug discovery: the role of analytical performance in ligand binding assays

Wätzig

Oltmann-Norden

Steinicke

et al. 2015

J Comput Aided Mol Des

View full text Add to dashboard Cite

Despite its importance and all the considerable efforts made, the progress in drug discovery is limited. One main reason for this is the partly questionable data quality. Models relating biological activity and structures and in silico predictions rely on precisely and accurately measured binding data. However, these data vary so strongly, such that only variations by orders of magnitude are considered as unreliable. This can certainly be improved considering the high analytical performance in pharmaceutical quality control. Thus the principles, properties and performances of biochemical and cell-based assays are revisited and evaluated. In the part of biochemical assays immunoassays, fluorescence assays, surface plasmon resonance, isothermal calorimetry, nuclear magnetic resonance and affinity capillary electrophoresis are discussed in details, in addition radiation-based ligand binding assays, mass spectrometry, atomic force microscopy and microscale thermophoresis are briefly evaluated. In addition, general sources of error, such as solvent, dilution, sample pretreatment and the quality of reagents and reference materials are discussed. Biochemical assays can be optimized to provide good accuracy and precision (e.g. percental relative standard deviation <10 %). Cell-based assays are often considered superior related to the biological significance, however, typically they cannot still be considered as really quantitative, in particular when results are compared over longer periods of time or between laboratories. A very careful choice of assays is therefore recommended. Strategies to further optimize assays are outlined, considering the evaluation and the decrease of the relevant error sources. Analytical performance and data quality are still advancing and will further advance the progress in drug development.

show abstract

“…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

confidence: 99%

Diet, behavior and immunity across the lifespan

Hale

Spencer

Conti

et al. 2015

Neuroscience & Biobehavioral Reviews

View full text Add to dashboard Cite

It is increasingly appreciated that perinatal events can set an organism on a life-long trajectory for either health or disease, resilience or risk. One early life variable that has proven critical for optimal development is the nutritional environment in which the organism develops. Extensive research has documented the effects of both undernutrition and overnutrition, with strong links evident for an increased risk for obesity and metabolic disorders, as well as adverse mental health outcomes. Recent work has highlighted a critical role of the immune system, in linking diet with long term health and behavioral outcomes. The present review will summarize the recent literature regarding the interactions of diet, immunity, and behavior.

show abstract

Design and validation of a homogeneous time-resolved fluorescence-based leptin receptor binding assay

Cited by 20 publications

References 35 publications

Endospanin 1 Determines the Balance of Leptin-Regulated Hypothalamic Functions

Endospanin 1 Determines the Balance of Leptin-Regulated Hypothalamic Functions

Data quality in drug discovery: the role of analytical performance in ligand binding assays

Diet, behavior and immunity across the lifespan

Contact Info

Product

Resources

About