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The human GABAB receptor-a member of the class C family of G-protein-coupled receptors (GPCRs)-mediates inhibitory neurotransmission and has been implicated in epilepsy, pain and addiction1. A unique GPCR that is known to require heterodimerization for function2-6, the GABAB receptor has two subunits, GABAB1 and GABAB2, that are structurally homologous but perform distinct and complementary functions. GABAB1 recognizes orthosteric ligands7,8, while GABAB2 couples with G proteins9-14. Each subunit is characterized by an extracellular Venus flytrap (VFT) module, a descending peptide linker, a seven-helix transmembrane domain and a cytoplasmic tail15. Although the VFT heterodimer structure has been resolved16, the structure of the full-length receptor and its transmembrane signalling mechanism remain unknown. Here we present a near full-length structure of the GABAB receptor, captured in an inactive state by cryo-electron microscopy. Our structure reveals several ligands that preassociate with the receptor, including two large endogenous phospholipids that are embedded within the transmembrane domains to maintain receptor integrity and modulate receptor function. We also identify a previously unknown heterodimer interface between transmembrane helices 3 and 5 of both subunits, which serves as a signature of the inactive conformation. A unique 'intersubunit latch' within this transmembrane interface maintains the inactive state, and its disruption leads to constitutive receptor activity.

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Classically, G-protein-coupled receptors (GPCRs) are thought to activate G protein from the plasma membrane and are subsequently desensitized by β-arrestin (β-arr). However, some GPCRs continue to signal through G protein from internalized compartments, mediated by a GPCR-G protein-β-arr 'megaplex'. Nevertheless, the molecular architecture of the megaplex remains unknown. Here, we present its cryo-electron microscopy structure, which shows simultaneous engagement of human G protein and bovine β-arr to the core and phosphorylated tail, respectively, of a single active human chimeric β2-adrenergic receptor with the C-terminal tail of the arginine vasopressin type 2 receptor (β2V2R). All three components adopt their canonical active conformations, suggesting that a single megaplex GPCR is capable of simultaneously activating G protein and β-arr. Our findings provide a structural basis for GPCR-mediated sustained internalized G protein signaling.

We study point-line configurations through the lens of projective geometry and matroid theory. Our focus is on their realization spaces, where we introduce the concepts of liftable and quasi-liftable configurations, exploring cases in which an n-tuple of collinear points can be lifted to a nondegenerate realization of a point-line configuration. We show that forest configurations are liftable and characterize the realization space of liftable configurations as the solution set of certain linear systems of equations. Moreover, we study the Zariski closure of the realization spaces of liftable and quasi-liftable configurations, known as matroid varieties, and establish their irreducibility. Additionally, we compute an irreducible decomposition for their corresponding circuit varieties. Applying these liftability properties, we present a procedure to generate some of the defining equations of the associated matroid varieties. As corollaries, we provide a geometric representation for the defining equations of two specific examples: the quadrilateral set and the 3 × 4 grid. While the polynomials for the latter were previously computed using specialized algorithms tailored for this configuration, the geometric interpretation of these generators was missing. We compute a minimal generating set for the corresponding ideals. [ABSTRACT FROM AUTHOR]

HCN1-4 channels are the molecular determinants of the If/Ih current that crucially regulates cardiac and neuronal cell excitability. HCN dysfunctions lead to sinoatrial block (HCN4), epilepsy (HCN1), and chronic pain (HCN2), widespread medical conditions awaiting subtype-specific treatments. Here, we address the problem by solving the cryo-EM structure of HCN4 in complex with ivabradine, to date the only HCN-specific drug on the market. Our data show ivabradine bound inside the open pore at 3 Å resolution. The structure unambiguously proves that Y507 and I511 on S6 are the molecular determinants of ivabradine binding to the inner cavity, while F510, pointing outside the pore, indirectly contributes to the block by controlling Y507. Cysteine 479, unique to the HCN selectivity filter (SF), accelerates the kinetics of block. Molecular dynamics simulations further reveal that ivabradine blocks the permeating ion inside the SF by electrostatic repulsion, a mechanism previously proposed for quaternary ammonium ions. [ABSTRACT FROM AUTHOR]

We study varieties associated to hypergraphs from the point of view of projective geometry and matroid theory. We describe their decompositions into matroid varieties, which may be reducible and can have arbitrary singularities by the Mnëv–Sturmfels universality theorem. We focus on various families of hypergraph varieties for which we explicitly compute an irredundant irreducible decomposition. Our main findings in this direction are three-fold: (1) we describe minimal matroids of such hypergraphs; (2) we prove that the varieties of these matroids are irreducible and their union is the hypergraph variety; and (3) we show that every such matroid is realizable over real numbers. As corollaries, we give conceptual decompositions of various, previously studied, varieties associated with graphs, hypergraphs, and adjacent minors of generic matrices. In particular, our decomposition strategy gives immediate matroid interpretations of the irreducible components of multiple families of varieties associated to conditional independence models in statistical theory and unravels their symmetric structures which hugely simplifies the computations. [ABSTRACT FROM AUTHOR]

Thyroglobulin must pass endoplasmic reticulum (ER) quality control to become secreted for thyroid hormone synthesis. Defective thyroglobulin, blocked in trafficking, can cause hypothyroidism. Thyroglobulin is a large protein (~2750 residues) spanning regions I–II–III plus a C‐terminal cholinesterase‐like domain. The cholinesterase‐like domain functions as an intramolecular chaperone for regions I–II–III, but the folding pathway leading to successful thyroglobulin trafficking remains largely unknown. Here, informed by the recent three‐dimensional structure of thyroglobulin as determined by cryo‐electron microscopy, we have bioengineered three novel classes of mutants yielding three entirely distinct quality control phenotypes. Specifically, upon expressing recombinant thyroglobulin, we find that first, mutations eliminating a disulfide bond enclosing a 200‐amino acid loop in region I have surprisingly little impact on the ability of thyroglobulin to fold to a secretion‐competent state. Next, we have identified a mutation on the surface of the cholinesterase‐like domain that has no discernible effect on regional folding yet affects contact between distinct regions and thereby triggers impairment in the trafficking of full‐length thyroglobulin. Finally, we have probed a conserved disulfide in the cholinesterase‐like domain that interferes dramatically with local folding, and this defect then impacts on global folding, blocking the entire thyroglobulin in the ER. These data highlight variants with distinct effects on ER quality control, inhibiting domain‐specific folding; folding via regional contact; neither; or both. [ABSTRACT FROM AUTHOR]

We study the equivariant Ehrhart theory of families of polytopes that are invariant under a non-trivial action of the group with order two. We study families of polytopes whose equivariant H^\ast-polynomial both succeed and fail to be effective, in particular, the symmetric edge polytopes of cycle graphs and the rational cross-polytope. The latter provides a counterexample to the effectiveness conjecture if the requirement that the vertices of the polytope have integral coordinates is loosened to allow rational coordinates. Moreover, we exhibit such a counterexample whose Ehrhart function has period one and coincides with the Ehrhart function of a lattice polytope. [ABSTRACT FROM AUTHOR]

Ryanodine receptors (RyRs) mediate the rapid release of calcium ([Ca.sup.2+]) from intracellular stores into the cytosol, which is essential for numerous cellular functions including excitation-contraction coupling in muscle. Lack of sufficient structural detail has impeded understanding of RyR gating and regulation. Here we report the closed-state structure of the 2.3-megadalton complex of the rabbit skeletal muscle type 1 RyR (RyR1), solved by single-particle electron cryomicroscopy at an overall resolution of 4.8 Å. We fitted a polyalanine-level model to all 3,757 ordered residues in each protomer, defining the transmembrane pore in unprecedented detail and placing all cytosolic domains as tertiary folds. The cytosolic assembly is built on an extended α-solenoid scaffold connecting key regulatory domains to the pore. The RyR1 pore architecture places it in the six-transmembrane ion channel superfamily. A unique domain inserted between the second and third transmembrane helices interacts intimately with paired EF-hands originating from the α-solenoid scaffold, suggesting a mechanism for channel gating by [Ca.sup.2+]., Ryanodine receptors (RyRs) are intracellular [Ca.sup.2+] release channels on the sarcoplasmic and endoplasmic reticula required for fundamental cellular functions in most tissues, including skeletal and cardiac muscle excitation-contraction coupling, synaptic [...]

A family of leucine-rich-repeat-containing G-protein-coupled receptors (LGRs) mediate diverse physiological responses when complexed with their cognate ligands. LGRs are present in all metazoan animals. In humans, the LGR ligands include glycoprotein hormones (GPHs) chorionic gonadotropin (hCG), luteinizing hormone, follicle-stimulating hormone (hFSH), and thyroid-stimulating hormone (hTSH). These hormones are aß heterodimers of cystine-knot protein chains. LGRs and their ligand chains have coevolved. Ancestral hormone homologs, present in both bilaterian animals and chordates, are identified as a2ß5. We have used single-wavelength anomalous diffraction and molecular replacement to determine structures of the a2ß5 hormone from Caenorhabditis elegans (Cea2ß5). Cea2ß5 is unglycosylated, as are many other a2ß5 hormones. Both Hsa2ß5, the human homolog of Cea2ß5, and hTSH activate the same receptor (hTSHR). Despite having little sequence similarity to vertebrate GPHs, apart from the cysteine patterns from core disulfide bridges, Cea2ß5 is generally similar in structure to these counterparts; however, its a2 and ß5 subunits are more symmetric as compared with a and ß of hCG and hFSH. This quasisymmetry suggests a hypothetical homodimeric antecedent of the a2ß5 and aß heterodimers. Known structures together with AlphaFold models from the sequences for other LGR ligands provide representatives for the molecular evolution of LGR ligands from early metazoans through the present-day GPHs. The experimental Cea2ß5 structure validates its AlphaFold model, and thus also that for Hsa2ß5; and interfacial characteristics in a model for the Hsa2ß5:hTSHR complex are similar to those found in an experimental hTSH:hTSHR structure. [ABSTRACT FROM AUTHOR]

We study the connection between probability distributions satisfying certain conditional independence (CI) constraints, and point and line arrangements in incidence geometry. To a family of CI statements, we associate a polynomial ideal whose algebraic invariants are encoded in a hypergraph. The primary decompositions of these ideals give a characterization of the distributions satisfying the original CI statements. Classically, these ideals are generated by 2-minors of a matrix of variables; however, in the presence of hidden variables, they contain higher degree minors. This leads to the study of the structure of determinantal hypergraph ideals whose decompositions can be understood in terms of point and line configurations in the projective space. [ABSTRACT FROM AUTHOR]

The primary functional units of the thyroid gland are follicles of various sizes comprised of a monolayer of epithelial cells (thyrocytes) surrounding an apical extracellular cavity known as the follicle lumen. In the normal thyroid gland, the follicle lumen is filled with secreted protein (referred to as colloid), comprised nearly exclusively of thyroglobulin with a half-life ranging from days to weeks. At the cellular boundary of the follicle lumen, secreted thyroglobulin becomes iodinated, resulting from the coordinated activities of enzymes localized to the thyrocyte apical plasma membrane. Thyroglobulin appearance in evolution is essentially synchronous with the appearance of the follicular architecture of the vertebrate thyroid gland. Thyroglobulin is the most highly expressed thyroid gene and represents the most abundantly expressed thyroid protein. Wildtype thyroglobulin protein is a large and complex glycoprotein that folds in the endoplasmic reticulum, leading to homodimerization and export via the classical secretory pathway to the follicle lumen. However, of the hundreds of human thyroglobulin genetic variants, most exhibit increased susceptibility to misfolding with defective export from the endoplasmic reticulum, triggering hypothyroidism as well as thyroidal endoplasmic reticulum stress. The human disease of hypothyroidism with defective thyroglobulin (either homozygous, or compound heterozygous) can be experimentally modeled in thyrocyte cell culture, or in whole animals, such as mice that are readily amenable to genetic manipulation. From a combination of approaches, it can be demonstrated that in the setting of thyroglobulin misfolding, thyrocytes under chronic continuous ER stress exhibit increased susceptibility to cell death, with interesting cell biological and pathophysiological consequences. [ABSTRACT FROM AUTHOR]

Tetratricopeptide repeat–containing Rab8b-interacting (TRIP8b) protein is a brain-specific subunit of Hyperpolarization-activated Cyclic Nucleotide-gated (HCN) channels, a class of voltage-gated channels modulated by cyclic nucleotides. While the interaction between TRIP8b and the cytosolic C terminus of the channel has been structurally described, the HCN:TRIP8b stoichiometry is less characterized. We employed single molecule mass photometry (MP) to image HCN4 particles purified in complex with TRIP8b. Our data show that four TRIP8b subunits are bound to the tetrameric HCN4 particle, confirming a 1:1 stoichiometry. [ABSTRACT FROM AUTHOR]