Evidence for leptin receptor isoforms heteromerization at the cell surface

Bacart, Johan; Leloire, Audrey; Levoye, Angélique; Froguel, Philippe; Jockers, Ralf; Couturier, Cyril

doi:10.1016/j.febslet.2010.03.033

Cited by 24 publications

(24 citation statements)

References 25 publications

(62 reference statements)

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“…This range from two soluble proteins, two transmembraneous ones, one transmembraneous, and one soluble, with interactions taking place in cytoplasm, nucleus, and cytoplasmic or internal membranes (Coulon et al, 2008; Guan et al, 2009; Bacart et al, 2010). Indeed BRET is able to monitor all kinds of interaction, however, certain concerns have to be taken into account when designing P2I2 BRET-based assays.…”

Section: For Which Kind Of Target Interaction Can the Bret Be Chosen?mentioning

confidence: 99%

Setting Up a Bioluminescence Resonance Energy Transfer High throughput Screening Assay to Search for Protein/Protein Interaction Inhibitors in Mammalian Cells

Couturier¹,

Deprez²

2012

Front. Endocrin.

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Each step of the cell life and its response or adaptation to its environment are mediated by a network of protein/protein interactions termed “interactome.” Our knowledge of this network keeps growing due to the development of sensitive techniques devoted to study these interactions. The bioluminescence resonance energy transfer (BRET) technique was primarily developed to allow the dynamic monitoring of protein/protein interactions (PPI) in living cells, and has widely been used to study receptor activation by intra- or extra-molecular conformational changes within receptors and activated complexes in mammal cells. Some interactions are described as crucial in human pathological processes, and a new class of drugs targeting them has recently emerged. The BRET method is well suited to identify inhibitors of PPI and here is described why and how to set up and optimize a high throughput screening assay based on BRET to search for such inhibitory compounds. The different parameters to take into account when developing such BRET assays in mammal cells are reviewed to give general guidelines: considerations on the targeted interaction, choice of BRET version, inducibility of the interaction, kinetic of the monitored interaction, and of the BRET reading, influence of substrate concentration, number of cells and medium composition used on the Z′ factor, and expected interferences from colored or fluorescent compounds.

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Section: For Which Kind Of Target Interaction Can the Bret Be Chosen?mentioning

confidence: 99%

Setting Up a Bioluminescence Resonance Energy Transfer High throughput Screening Assay to Search for Protein/Protein Interaction Inhibitors in Mammalian Cells

Couturier¹,

Deprez²

2012

Front. Endocrin.

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“…However, although there is considerable supporting evidence for the expression of alternative mature RNA transcripts, even when these transcripts are likely to encode protein sequences with unusual features (Yura et al 2006; Tress et al 2007; Melamud and Moult 2009), it has proved more difficult to demonstrate the existence of alternative protein isoforms at a global level. Much of the evidence for the translation of alternative splice variants as stable proteins has come from individual experiments (Guo et al 2003; Bacart et al 2010; Wang et al 2011). …”

Section: Introductionmentioning

confidence: 99%

Comparative Proteomics Reveals a Significant Bias Toward Alternative Protein Isoforms with Conserved Structure and Function

Ezkurdia

Pozo

Frankish

et al. 2012

Molecular Biology and Evolution

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Advances in high-throughput mass spectrometry are making proteomics an increasingly important tool in genome annotation projects. Peptides detected in mass spectrometry experiments can be used to validate gene models and verify the translation of putative coding sequences (CDSs). Here, we have identified peptides that cover 35% of the genes annotated by the GENCODE consortium for the human genome as part of a comprehensive analysis of experimental spectra from two large publicly available mass spectrometry databases. We detected the translation to protein of “novel” and “putative” protein-coding transcripts as well as transcripts annotated as pseudogenes and nonsense-mediated decay targets.We provide a detailed overview of the population of alternatively spliced protein isoforms that are detectable by peptide identification methods. We found that 150 genes expressed multiple alternative protein isoforms. This constitutes the largest set of reliably confirmed alternatively spliced proteins yet discovered. Three groups of genes were highly overrepresented. We detected alternative isoforms for 10 of the 25 possible heterogeneous nuclear ribonucleoproteins, proteins with a key role in the splicing process. Alternative isoforms generated from interchangeable homologous exons and from short indels were also significantly enriched, both in human experiments and in parallel analyses of mouse and Drosophila proteomics experiments. Our results show that a surprisingly high proportion (almost 25%) of the detected alternative isoforms are only subtly different from their constitutive counterparts.Many of the alternative splicing events that give rise to these alternative isoforms are conserved in mouse. It was striking that very few of these conserved splicing events broke Pfam functional domains or would damage globular protein structures. This evidence of a strong bias toward subtle differences in CDS and likely conserved cellular function and structure is remarkable and strongly suggests that the translation of alternative transcripts may be subject to selective constraints.

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“…On the membrane, however, the LR can assemble as preformed dimers or oligomers, as evidenced by a high basal signal in the absence of leptin in both fluorescence resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET) between differently tagged LRs and by co-immunoprecipitation of tagged LRs (Nakashima et al 1997, White & Tartaglia 1999, Couturier & Jockers 2003, Biener et al 2005. Different isoforms of the LR can heterodimerize with each other both in the presence and absence of leptin (White & Tartaglia 1999, Bacart et al 2010. Leptin treatment induces an increase in BRET and FRET signals, which are probably a consequence of reorganization within the pre-formed complexes and/or de novo oligomerization (Couturier & Jockers 2003, Biener et al 2005.…”

Section: Disulfide Network and Glycosylation Of The Ecdmentioning

confidence: 99%

20 YEARS OF LEPTIN: Insights into signaling assemblies of the leptin receptor

Peelman¹,

Zabeau²,

Moharana³

et al. 2014

Journal of Endocrinology

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Leptin plays a central role in the control of body weight and energy homeostasis, but is a pleiotropic cytokine with activities on many peripheral cell types. In this review, we discuss the interaction of leptin with its receptor, and focus on the structural and mechanistic aspects of the extracellular aspects of leptin receptor (LR) activation. We provide an extensive overview of all structural information that has been obtained for leptin and its receptor via X-ray crystallography, electron microscopy, small-angle X-ray scattering, homology modeling, and mutagenesis studies. The available knowledge is integrated into putative models toward a recapitulation of the LR activation mechanism.

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Evidence for leptin receptor isoforms heteromerization at the cell surface

Cited by 24 publications

References 25 publications

Setting Up a Bioluminescence Resonance Energy Transfer High throughput Screening Assay to Search for Protein/Protein Interaction Inhibitors in Mammalian Cells

Setting Up a Bioluminescence Resonance Energy Transfer High throughput Screening Assay to Search for Protein/Protein Interaction Inhibitors in Mammalian Cells

Comparative Proteomics Reveals a Significant Bias Toward Alternative Protein Isoforms with Conserved Structure and Function

20 YEARS OF LEPTIN: Insights into signaling assemblies of the leptin receptor

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