Visualization of ligand-induced transmembrane signaling in the full-length human insulin receptor

Gutmann, Theresia; Kim, Kelly H.; Grzybek, Michał; Walz, Thomas; Coskun, Ünal

doi:10.1083/jcb.201711047

Cited by 104 publications

(130 citation statements)

References 41 publications

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“…S4, S5). Our 3D reconstruction confirmed the T-like structure as seen in the insulin-bound full-length IR at low resolution by negative stain EM (Gutmann et al, 2018). Features of the compact head containing L1, CR, and L2 domains appear better defined than the fibronectin stalks, which exhibit more flexibility (cp.…”

Section: Single-particle Cryo-em Analysis Of the Ir-ecdsupporting

confidence: 71%

“…Insulin binding converts the receptor ectodomain into a T-like shape that draws the membraneproximal fibronectin domains closer together, enabling transmembrane signaling (Gutmann et al, 2018). Due to the low resolution of the negative-stain 2D class averages, no structural information about the location and number of bound insulin molecules could be obtained.…”

Section: Introductionmentioning

confidence: 99%

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Cryo-EM structure of the complete and ligand-saturated insulin receptor ectodomain

Gutmann

Schäfer

Poojari³

et al. 2019

Preprint

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Glucose homeostasis and growth essentially depend on the peptide hormone insulin engaging its receptor. Despite biochemical and structural advances, a fundamental contradiction has persisted in the current understanding of insulin ligand-receptor interactions. While biochemistry predicts two distinct insulin binding sites, 1 and 2, recent structural analyses have only resolved site 1.Using a combined approach of cryo-EM and atomistic molecular dynamics simulation, we determined the structure of the entire dimeric insulin receptor ectodomain saturated with four insulin molecules. Complementing the previously described insulin-site 1 interaction, we present the first view of insulin bound to the discrete insulin receptor site 2. Insulin binding stabilizes the receptor ectodomain in a T-shaped conformation wherein the membrane-proximal domains converge and contact each other. These findings expand the current models of insulin binding to its receptor and of its regulation. In summary, we provide the structural basis enabling a comprehensive description of ligand-receptor interactions that ultimately will inform new approaches to structure-based drug design. In briefA cryo-EM structure of the complete insulin receptor ectodomain saturated with four insulin ligands is reported. The structural model of the insulin-insulin receptor complex adopts a T-shaped conformation, reveals two additional insulin-binding sites potentially involved in the initial interaction of insulin with its receptor, and resolves the membrane proximal region.

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Section: Single-particle Cryo-em Analysis Of the Ir-ecdsupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Cryo-EM structure of the complete and ligand-saturated insulin receptor ectodomain

Gutmann

Schäfer

Poojari³

et al. 2019

Preprint

Self Cite

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“…Two lines of evidence support the view that the Λ-shaped structure represents that adopted by the ectodomain in the context of a membrane-embedded holo receptor. First, recent negative-stain electron microscopy imaging of holo insulin receptor embedded in lipid nanodiscs show, at least qualitatively, that the Λ-shaped assembly concurs with that observed in the nanodisc context 92 (Figure 10). Second, crystallographic analysis of the isolated and growth factor-free IGF-1R ectodomain reveals it to have a similar shape and domain assembly to that of the insulin receptor, albeit that the IGF-1R construct employed included the attachment of a pair of antibody F v modules.…”

Section: Position B24mentioning

confidence: 58%

“…Negative‐stain transmission electron microscopy images of holo insulin receptor embedded in nanodiscs . The left‐hand panel is an image of the receptor in an insulin‐free state, the right‐hand panel is an image of the receptor in an insulin‐bound state, with the panels underneath each offering an interpretation of the respective images above.…”

Section: Interaction Of the Aromatic Triplet With The Receptor's Primmentioning

confidence: 99%

“…The schematic is based on PDB entry 4ZXB 48 FIGURE 10 Negative-stain transmission electron microscopy images of holo insulin receptor embedded in nanodiscs. 92 The left-hand panel is an image of the receptor in an insulin-free state, the right-hand panel is an image of the receptor in an insulin-bound state, with the panels underneath each offering an interpretation of the respective images above. The correspondence of the Λ-shaped particle in the left-hand panel to those of that of the crystal structure of the isolated ectodomain is apparent.…”

Section: Position B24mentioning

confidence: 99%

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A thing of beauty: Structure and function of insulin's “aromatic triplet”

Weiss

Lawrence

2018

Diabetes Obesity Metabolism

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The classical crystal structure of insulin was determined in 1969 by D.C. Hodgkin et al. following a 35-year program of research. This structure depicted a hexamer remarkable for its self-assembly as a zinc-coordinated trimer of dimer. Prominent at the dimer interface was an "aromatic triplet" of conserved residues at consecutive positions in the B chain: Phe , Phe and Tyr . The elegance of this interface inspired the Oxford team to poetry: "A thing of beauty is a joy forever" (John Keats as quoted by Blundell, T.L., et al. Advances in Protein Chemistry 26:279-286 [1972]). Here, we revisit this aromatic triplet in light of recent advances in the structural biology of insulin bound as a monomer to fragments of the insulin receptor. Such co-crystal structures have defined how these side chains pack at the primary hormone-binding surface of the receptor ectodomain. On receptor binding, the B-chain β-strand (residues B24-B28) containing the aromatic triplet detaches from the α-helical core of the hormone. Whereas Tyr lies at the periphery of the receptor interface and may functionally be replaced by a diverse set of substitutions, Phe and Phe engage invariant elements of receptor domains L1 and αCT. These critical contacts were anticipated by the discovery of diabetes-associated mutations at these positions by Donald Steiner et al. at the University of Chicago. Conservation of Phe , Phe and Tyr among vertebrate insulins reflects the striking confluence of structure-based evolutionary constraints: foldability, protective self-assembly and hormonal activity.

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Cryo‐Electron Microscopy in Drug Discovery and Development: Opportunities and Challenges

Gomez‐Llorente

Hong

Scapin

2021

Burger's Medicinal Chemistry and Drug Discovery

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While utilization of structural information has been a staple technology in drug discovery for many years, electron microscopy creates opportunities that have been thus far out of reach for the conventional techniques, like the visualization of very large molecular complexes in solution or obtaining high‐resolution structural data for small molecules from microcrystals. These opportunities come at a cost: hardware is extremely expensive, software is ever‐changing, and constantly challenged by new developments in the hardware and requests by the users, and IT departments are faced with significant requirements for storage and computational power. Nevertheless, the establishment of centralized facilities, commercial consortia, and cooperative research centers is well positioned to provide access to Cryo‐EM to an expanded base of researchers, from academia as well as from industry. Understanding of the requirements, advantages, and limitations of this technique becomes then necessary to ensure correct decision‐making processes and resource allocation when undertaking a structure‐guided drug design project.

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Visualization of ligand-induced transmembrane signaling in the full-length human insulin receptor

Cited by 104 publications

References 41 publications

Cryo-EM structure of the complete and ligand-saturated insulin receptor ectodomain

Cryo-EM structure of the complete and ligand-saturated insulin receptor ectodomain

A thing of beauty: Structure and function of insulin's “aromatic triplet”

Cryo‐Electron Microscopy in Drug Discovery and Development: Opportunities and Challenges

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