Biased signaling due to oligomerization of the G protein-coupled platelet-activating factor receptor

Liu, Junke; Tang, Hengmin; Xu, Chanjuan; Zhou, Shengnan; Zhu, Xunying; Li, Yuanyuan; Prézeau, Laurent; Xu, Tao; Pin, Jean‐Philippe; Rondard, Philippe; Ji, Wei; Liu, Jianfeng

doi:10.1038/s41467-022-34056-4

Cited by 32 publications

(25 citation statements)

References 110 publications

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“…The type of density-dependent affinity enhancement identified here for Ndc80-CENP-T binding can facilitate biogenesis of structures that rely on strong site-specific bonds (38), as well as modulate the activity of more transient oligomers and clusters, such as involved in signaling activity (39, 40), and chromatin-associated hubs (41). Thus, emergent behaviors involving slow binding-site maturation and density-induced acceleration could assist biomolecular self-organization and may facilitate engineering of self-assembling nanoparticles for medical and technological applications.…”

Section: Discussionmentioning

confidence: 99%

Binding Site Maturation Modulated by Molecular Density Underlies Ndc80 Binding to Kinetochore Receptor CENP-T

Tarasovetc,

Sissoko,

Maiorov

et al. 2024

Preprint

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Formation of macromolecular cellular structures relies on recruitment of multiple proteins, requiring the precisely controlled pairwise binding interactions. At human kinetochores, our recent work found that the high molecular density environment enables strong bonding between the Ndc80 complex and its two binding sites at the CENP-T receptor. However, the mechanistic basis for this unusual density-dependent facilitation remains unknown. Here, using quantitative single-molecule approaches, we reveal two distinct mechanisms that drive preferential recruitment of the Ndc80 complex to higher-order structures of CENP-T, as opposed to CENP-T monomers. First, the Ndc80 binding sites within the disordered tail of the CENP-T mature over time, leading to a stronger grip on the Spc24/25 heads of the Ndc80 complexes. Second, the maturation of Ndc80 binding sites is accelerated when CENP-T molecules are clustered in close proximity. The rates of the clustering-induced maturation are remarkably different for two binding sites within CENP-T, correlating with different interfaces formed by the corresponding CENP-T sequences as they wrap around the Spc24/25 heads. The differential clustering-dependent regulation of these sites is preserved in dividing human cells, suggesting a distinct regulatory entry point to control kinetochore-microtubule interactions. The tunable acceleration of slowly maturing binding sites by a high molecular-density environment may represent a fundamental physicochemical mechanism to assist the assembly of mitotic kinetochores and other macromolecular structures.

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Section: Discussionmentioning

confidence: 99%

Binding Site Maturation Modulated by Molecular Density Underlies Ndc80 Binding to Kinetochore Receptor CENP-T

Tarasovetc,

Sissoko,

Maiorov

et al. 2024

Preprint

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“…Site-directed mutations in the plasmids were generated using the QuikChange mutagenesis protocol (Agilent Technologies). The probes (full length of EGFP) were fused to the CTD of GLP-1R and MOR, respectively, with AflII restriction site as a linker 84 . The plasmids encoding human βarr1, βarr2, GRK2, GRK3, GRK5, and GRK6 in pcDNA3.1 were tagged with Flag-TST (provided by ARPEGE platform, IGF).…”

Section: Methodsmentioning

confidence: 99%

Multi-faceted roles of β-arrestins in G protein-coupled receptors endocytosis

Liu,

Xue,

Ravier

et al. 2024

Preprint

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Internalization plays a crucial role in regulating the density of cell surface receptors and has been demonstrated to regulate intracellular signaling. Dysregulation of this process has been implicated in various diseases. The vast majority of GPCRs were considered to adopt one way for internalization. We challenged this conventional view by showing that multiple pathways converge to regulate the internalization of a specific receptor, based on an unparalleled characterization of 60 GPCR internalization profiles, both in the absence and presence of individual βarrestins (βarrs). Furthermore, we revealed the internalization mechanism of the glucagon-like peptide-1 receptor (GLP-1R), a class B GPCR pivotal in promoting insulin secretion from pancreatic beta cells to maintain glucose homeostasis. GLP-1R can undergo agonist-induced internalization without βarrs, but can recruit and form stable complexes with βarrs. We found that GLP-1R recruits clathrin adaptor protein-2 for agonist-induced internalization in both βarr-dependent and -independent manners. These results provide a valuable resource for GPCR signaling and reveal the plasticity of different GPCRs to employ or not βarrs in the clathrin-mediated internalization.

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“…The ability of PSC to quantify the number of molecules in a molecular complex can reveal important information, regarding not only the size of the complex but also the role played by oligomerization in biological function. Liu et al 23 used photobleaching step analysis to assess the importance of oligomerization of the platelet-activation factor receptor (PAFR), a GPCR involved in platelet aggregation and inflammatory responses, in the particular signaling pathway initiated by the receptor. The study reports the formation of PAFR dimers and oligomers in transfected cell lines.…”

Section: Protein Colocalization For Detection Of Protein− Protein Int...mentioning

confidence: 99%

Fluorescence-Based Detection of Proteins and Their Interactions in Live Cells

Stoneman

Raicu

2023

J. Phys. Chem. B

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Recent advances in fluorescence-based microscopy techniques, such as single molecule fluorescence, Forster resonance energy transfer (FRET), fluorescence intensity fluctuations analysis, and super-resolution microscopy have expanded our ability to study proteins in greater detail within their native cellular environment and to investigate the roles that protein interactions play in biological functions, such as inter-and intracellular signaling and cargo transport. In this Perspective, we provide an up-to-date overview of the current state of the art in fluorescencebased detection of proteins and their interactions in living cells with an emphasis on recent developments that have facilitated the characterization of the spatial and temporal organization of proteins into oligomeric complexes in the presence and absence of natural and artificial ligands. Further advancements in this field will only deepen our understanding of the underlying mechanisms of biological processes and help develop new therapeutic targets.

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Biased signaling due to oligomerization of the G protein-coupled platelet-activating factor receptor

Cited by 32 publications

References 110 publications

Binding Site Maturation Modulated by Molecular Density Underlies Ndc80 Binding to Kinetochore Receptor CENP-T

Binding Site Maturation Modulated by Molecular Density Underlies Ndc80 Binding to Kinetochore Receptor CENP-T

Multi-faceted roles of β-arrestins in G protein-coupled receptors endocytosis

Fluorescence-Based Detection of Proteins and Their Interactions in Live Cells

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