المؤلفون: Kasaragod, Vikram Babu, Malinauskas, Tomas, Wahid, Ayla A, Lengyel, Judith, Knoflach, Frederic, Hardwick, Steven W, Jones, Charlotte F, Chen, Wan-Na, Lucas, Xavier, El Omari, Kamel, Chirgadze, Dimitri Y, Aricescu, A Radu, Cecere, Giuseppe, Hernandez, Maria-Clemencia, Miller, Paul S

مصطلحات موضوعية: Receptors, GABA-A, gamma-Aminobutyric Acid, Isoxazoles

الوصف: Acknowledgements: We acknowledge L. Cooper for training in cryo-EM grid preparation and for performing grid clipping; J. Stayaert (Vrije Universiteit Brussel) for kindly providing MbF3; M. Reutlinger and J. Benz for scientific discussions early in the project; V. Graf for electrophysiology support; and M. Karg, M. Fogetta and M. Siegrist for molecular biology support. This work was supported by a BBSRC project grant, BB/M024709/1 (P.S.M.), the Department of Pharmacology new lab start-up fund, and the University of Cambridge Isaac Newton & Wellcome Trust Institutional Strategic Support Fund, Academy of Medical Sciences Springboard Award, SBF004\1074 (P.S.M.), and funding from F. Hoffmann-La Roche Ltd. The cryo-EM facility receives funding from the Wellcome Trust, 206171/Z/17/Z; 202905/Z/16/Z (S.W.H. and D.Y.C.) and University of Cambridge. ; α5 subunit-containing γ-aminobutyric acid type A (GABAA) receptors represent a promising drug target for neurological and neuropsychiatric disorders. Altered expression and function contributes to neurodevelopmental disorders such as Dup15q and Angelman syndromes, developmental epilepsy and autism. Effective drug action without side effects is dependent on both α5-subtype selectivity and the strength of the positive or negative allosteric modulation (PAM or NAM). Here we solve structures of drugs bound to the α5 subunit. These define the molecular basis of binding and α5 selectivity of the β-carboline, methyl 6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM), type II benzodiazepine NAMs, and a series of isoxazole NAMs and PAMs. For the isoxazole series, each molecule appears as an 'upper' and 'lower' moiety in the pocket. Structural data and radioligand binding data reveal a positional displacement of the upper moiety containing the isoxazole between the NAMs and PAMs. Using a hybrid molecule we directly measure the functional contribution of the upper moiety to NAM versus PAM activity. Overall, these structures provide a framework by which to understand distinct ...

وصف الملف: text/xml; application/pdf; application/zip

العلاقة: https://www.repository.cam.ac.uk/handle/1810/362300Test

الإتاحة: https://www.repository.cam.ac.uk/handle/1810/362300Test

المؤلفون: Altemose, Nicolas, Noor, Nudrat, Bitoun, Emmanuelle, Tumian, Afidalina, Imbeault, Michael, Chapman, J Ross, Aricescu, A Radu, Myers, Simon R

مصطلحات موضوعية: Biochemistry and Cell Biology, Bioinformatics and Computational Biology, Biological Sciences, Genetics, Human Genome, Biotechnology, 1.1 Normal biological development and functioning, Underpinning research, Generic health relevance, Binding Sites, Chromosome Mapping, DNA, HEK293 Cells, Histone-Lysine N-Methyltransferase, Homologous Recombination, Humans, Meiosis, Protein Binding, Protein Multimerization, KRAB, PRDM9, chromosomes, evolutionary biology, genes, genomics, human, meiosis, recombination, transposable elements, zinc finger protein, Biological sciences, Biomedical and clinical sciences, Health sciences

الوصف: PRDM9 binding localizes almost all meiotic recombination sites in humans and mice. However, most PRDM9-bound loci do not become recombination hotspots. To explore factors that affect binding and subsequent recombination outcomes, we mapped human PRDM9 binding sites in a transfected human cell line and measured PRDM9-induced histone modifications. These data reveal varied DNA-binding modalities of PRDM9. We also find that human PRDM9 frequently binds promoters, despite their low recombination rates, and it can activate expression of a small number of genes including CTCFL and VCX. Furthermore, we identify specific sequence motifs that predict consistent, localized meiotic recombination suppression around a subset of PRDM9 binding sites. These motifs strongly associate with KRAB-ZNF protein binding, TRIM28 recruitment, and specific histone modifications. Finally, we demonstrate that, in addition to binding DNA, PRDM9's zinc fingers also mediate its multimerization, and we show that a pair of highly diverged alleles preferentially form homo-multimers.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/0rb6f7fcTest

المؤلفون: Doody, Karen M, Stanford, Stephanie M, Sacchetti, Cristiano, Svensson, Mattias ND, Coles, Charlotte H, Mitakidis, Nikolaos, Kiosses, William B, Bartok, Beatrix, Fos, Camille, Cory, Esther, Sah, Robert L, Liu-Bryan, Ru, Boyle, David L, Arnett, Heather A, Mustelin, Tomas, Corr, Maripat, Esko, Jeffrey D, Tremblay, Michel L, Firestein, Gary S, Aricescu, A Radu, Bottini, Nunzio

المصدر: Science Translational Medicine. 7(288)

مصطلحات موضوعية: Medical Biotechnology, Biomedical and Clinical Sciences, Autoimmune Disease, Arthritis, Rheumatoid Arthritis, Aetiology, 2.1 Biological and endogenous factors, Musculoskeletal, Inflammatory and immune system, Animals, Antirheumatic Agents, Arthritis, Rheumatoid, Cell Adhesion, Cell Movement, Chondroitin Sulfate Proteoglycans, Cytoskeletal Proteins, Disease Models, Animal, HEK293 Cells, Heparin, Humans, Mice, Knockout, Molecular Targeted Therapy, Phosphorylation, Proteoglycans, Receptor-Like Protein Tyrosine Phosphatases, Class 2, Severity of Illness Index, Signal Transduction, Syndecan-4, Synovial Membrane, Time Factors, Transfection, Biological Sciences, Medical and Health Sciences, Medical biotechnology, Biomedical engineering

الوصف: Despite the availability of several therapies for rheumatoid arthritis (RA) that target the immune system, a large number of RA patients fail to achieve remission. Joint-lining cells, called fibroblast-like synoviocytes (FLS), become activated during RA and mediate joint inflammation and destruction of cartilage and bone. We identify RPTPσ, a transmembrane tyrosine phosphatase, as a therapeutic target for FLS-directed therapy. RPTPσ is reciprocally regulated by interactions with chondroitin sulfate or heparan sulfate containing extracellular proteoglycans in a mechanism called the proteoglycan switch. We show that the proteoglycan switch regulates FLS function. Incubation of FLS with a proteoglycan-binding RPTPσ decoy protein inhibited cell invasiveness and attachment to cartilage by disrupting a constitutive interaction between RPTPσ and the heparan sulfate proteoglycan syndecan-4. RPTPσ mediated the effect of proteoglycans on FLS signaling by regulating the phosphorylation and cytoskeletal localization of ezrin. Furthermore, administration of the RPTPσ decoy protein ameliorated in vivo human FLS invasiveness and arthritis severity in the K/BxN serum transfer model of RA. Our data demonstrate that FLS are regulated by an RPTPσ-dependent proteoglycan switch in vivo, which can be targeted for RA therapy. We envision that therapies targeting the proteoglycan switch or its intracellular pathway in FLS could be effective as a monotherapy or in combination with currently available immune-targeted agents to improve control of disease activity in RA patients.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/81n774zgTest

المؤلفون: Piot, Laura, Heroven, Christina, Bossi, Simon, Zamith, Joseph, Malinauskas, Tomas, Johnson, Chris, Wennagel, Doris, Stroebel, David, Charrier, Cécile, Aricescu, A. Radu, Mony, Laetitia, Paoletti, Pierre

المصدر: Science; 12/22/2023, Vol. 382 Issue 6677, p1389-1394, 6p, 4 Graphs

المؤلفون: Salzer, Ralf, Clark, Jordan J, Vaysburd, Marina, Chang, Veronica T, Albecka, Anna, Kiss, Leo, Sharma, Parul, Gonzalez Llamazares, Andres, Kipar, Anja, Hiscox, Julian A, Owen, Andrew, Aricescu, A Radu, Stewart, James P, James, Leo C, Löwe, Jan

المصدر: Salzer, Ralf; Clark, Jordan J; Vaysburd, Marina; Chang, Veronica T; Albecka, Anna; Kiss, Leo; Sharma, Parul; Gonzalez Llamazares, Andres; Kipar, Anja; Hiscox, Julian A; Owen, Andrew; Aricescu, A Radu; Stewart, James P; James, Leo C; Löwe, Jan (2021). Single‐dose immunisation with a multimerised SARS‐CoV‐2 receptor binding domain (RBD) induces an enhanced and protective response in mice. FEBS letters, 595(18):2323-2340.

مصطلحات موضوعية: Institute of Veterinary Pathology, 570 Life sciences, biology, Cell Biology, Genetics, Molecular Biology, Biochemistry, Structural Biology, Biophysics

الوصف: The COVID-19 pandemic, caused by the SARS-CoV-2 coronavirus, has triggered a worldwide health emergency. Here, we show that ferritin-like Dps from hyperthermophilic Sulfolobus islandicus, covalently coupled with SARS-CoV-2 antigens via the SpyCatcher system, forms stable multivalent dodecameric vaccine nanoparticles that remain intact even after lyophilisation. Immunisation experiments in mice demonstrated that the SARS-CoV-2 receptor binding domain (RBD) coupled to Dps (RBD-S-Dps) elicited a higher antibody titre and an enhanced neutralising antibody response compared to monomeric RBD. A single immunisation with RBD-S-Dps completely protected hACE2-expressing mice from serious illness and led to viral clearance from the lungs upon SARS-CoV-2 infection. Our data highlight that multimerised SARS-CoV-2 subunit vaccines are a highly efficacious modality, particularly when combined with an ultra-stable scaffold.

وصف الملف: application/pdf

العلاقة: https://www.zora.uzh.ch/id/eprint/205868/1/Kipar_2021_August.pdfTest; urn:issn:0014-5793

الإتاحة: https://doi.org/10.5167/uzh-20586810.1002/1873-3468.14171Test
https://www.zora.uzh.ch/id/eprint/205868Test/
https://www.zora.uzh.ch/id/eprint/205868/1/Kipar_2021_August.pdfTest

المؤلفون: Robinson, Ross A., Griffiths, Samuel C., van de Haar, Lieke L., Malinauskas, Tomas, van Battum, Eljo Y., Zelina, Pavol, Schwab, Rebekka A., Karia, Dimple, Malinauskaite, Lina, Brignani, Sara, van den Munkhof, Marleen H., Düdükcü, Özge, De Ruiter, Anna A., Van den Heuvel, Dianne M.A., Bishop, Benjamin, Elegheert, Jonathan, Aricescu, A. Radu, Pasterkamp, R. Jeroen, Siebold, Christian

المساهمون: TN groep Pasterkamp, Projectafdeling ALS, Translational Neuroscience, Brain, Regenerative Medicine and Stem Cells

مصطلحات موضوعية: axon regeneration, cell migration, cell surface receptors, complex structure, morphogen signaling, Neogenin, Netrin, protein-protein interactions, repulsive guidance molecule, signal transduction, Bioengineering, Medicine (miscellaneous), Biomedical Engineering, General Biochemistry,Genetics and Molecular Biology

الوصف: During cell migration or differentiation, cell surface receptors are simultaneously exposed to different ligands. However, it is often unclear how these extracellular signals are integrated. Neogenin (NEO1) acts as an attractive guidance receptor when the Netrin-1 (NET1) ligand binds, but it mediates repulsion via repulsive guidance molecule (RGM) ligands. Here, we show that signal integration occurs through the formation of a ternary NEO1-NET1-RGM complex, which triggers reciprocal silencing of downstream signaling. Our NEO1-NET1-RGM structures reveal a “trimer-of-trimers” super-assembly, which exists in the cell membrane. Super-assembly formation results in inhibition of RGMA-NEO1-mediated growth cone collapse and RGMA- or NET1-NEO1-mediated neuron migration, by preventing formation of signaling-compatible RGM-NEO1 complexes and NET1-induced NEO1 ectodomain clustering. These results illustrate how simultaneous binding of ligands with opposing functions, to a single receptor, does not lead to competition for binding, but to formation of a super-complex that diminishes their functional outputs.

وصف الملف: application/pdf

العلاقة: https://dspace.library.uu.nl/handle/1874/442714Test

الإتاحة: https://dspace.library.uu.nl/handle/1874/442714Test

المؤلفون: Robinson, Ross, Griffiths, Samuel, van de Haar, Lieke, Malinauskas, Tomas, Van Battum, Eljo, Zelina, Pavol, Schwab, Rebekka, Karia, Dimple, Malinauskaite, Lina, Brignani, Sara, van den Munkhof, Marleen, Düdükcü, Özge, De Ruiter, Anna, Van den Heuvel, Dianne M.A., Bishop, Benjamin, Elegheert, Jonathan, Aricescu, A. Radu, Pasterkamp, R. Jeroen, Siebold, Christian

المساهمون: Interdisciplinary Institute for Neuroscience Bordeaux (IINS), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS)

المصدر: ISSN: 0092-8674.

مصطلحات موضوعية: [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology

الوصف: International audience

العلاقة: hal-03396256; https://hal.archives-ouvertes.fr/hal-03396256Test; https://hal.archives-ouvertes.fr/hal-03396256/documentTest; https://hal.archives-ouvertes.fr/hal-03396256/file/2021_Cell_Robinson_NEO-RGM-NET_merged.pdfTest

الإتاحة: https://doi.org/10.1016/j.cell.2021.02.045Test
https://hal.archives-ouvertes.fr/hal-03396256Test
https://hal.archives-ouvertes.fr/hal-03396256/documentTest
https://hal.archives-ouvertes.fr/hal-03396256/file/2021_Cell_Robinson_NEO-RGM-NET_merged.pdfTest

المؤلفون: Elegheert, Jonathan, Cvetkovska, Vedrana, Clayton, Amber J., Heroven, Christina, Vennekens, Kristel M., Smukowski, Samuel N., Regan, Michael C., Jia, Wanyi, Smith, Alexandra C., Furukawa, Hiro, Savas, Jeffrey N., de Wit, Joris, Begbie, Jo, Craig, Ann Marie, Aricescu, A. Radu

المصدر: Neuron ; volume 109, issue 1, page 189-190 ; ISSN 0896-6273

مصطلحات موضوعية: General Neuroscience

الإتاحة: https://doi.org/10.1016/j.neuron.2020.12.006Test
https://api.elsevier.com/content/article/PII:S089662732030965X?httpAccept=text/xmlTest
https://api.elsevier.com/content/article/PII:S089662732030965X?httpAccept=text/plainTest

المؤلفون: Kasaragod, Vikram Babu, Malinauskas, Tomas, Wahid, Ayla A., Lengyel, Judith, Knoflach, Frederic, Hardwick, Steven W., Jones, Charlotte F., Chen, Wan-Na, Lucas, Xavier, El Omari, Kamel, Chirgadze, Dimitri Y., Aricescu, A. Radu, Cecere, Giuseppe, Hernandez, Maria-Clemencia, Miller, Paul S.

المصدر: Nature Structural and Molecular Biology; December 2023, Vol. 30 Issue: 12 p1936-1946, 11p

مستخلص: α5 subunit-containing γ-aminobutyric acid type A (GABAA) receptors represent a promising drug target for neurological and neuropsychiatric disorders. Altered expression and function contributes to neurodevelopmental disorders such as Dup15q and Angelman syndromes, developmental epilepsy and autism. Effective drug action without side effects is dependent on both α5-subtype selectivity and the strength of the positive or negative allosteric modulation (PAM or NAM). Here we solve structures of drugs bound to the α5 subunit. These define the molecular basis of binding and α5 selectivity of the β-carboline, methyl 6,7-dimethoxy-4-ethyl-β-carboline-3-carboxylate (DMCM), type II benzodiazepine NAMs, and a series of isoxazole NAMs and PAMs. For the isoxazole series, each molecule appears as an ‘upper’ and ‘lower’ moiety in the pocket. Structural data and radioligand binding data reveal a positional displacement of the upper moiety containing the isoxazole between the NAMs and PAMs. Using a hybrid molecule we directly measure the functional contribution of the upper moiety to NAM versus PAM activity. Overall, these structures provide a framework by which to understand distinct modulator binding modes and their basis of α5-subtype selectivity, appreciate structure–activity relationships, and empower future structure-based drug design campaigns.

المؤلفون: Aricescu, Alexandru Radu

مصطلحات موضوعية: 616, Biochemistry

الوصف: Receptor protein tyrosine phosphatases (RPTPs) represent a major research focus in developmental biology, oncology and immunology. Specifically, cell adhesion molecule-like RPTPs, including CRYPα, have recently emerged as key regulators of nervous system development, being involved in axonal growth and guidance, and nerve repair. However, very little is known about their signal transduction mechanisms since the identity of various interacting molecules is far from being completely elucidated. In this thesis I demonstrate that CRYPα has a novel, heparin-binding activity and that its interaction with the retinal basal lamina is mediated by the heparan sulphate chains of extracellular matrix proteoglycans. Using molecular modelling and site-directed mutagenesis, I have mapped the heparin/heparan sulphate binding site in the first immunoglobulin-like domain of CRYPα. I also describe the first identification of heterotypic ligands for a type II neural RPTP, namely the secreted isoforms of agrin and collagen XVIII. Given the broad functional spectrum of these molecules, a novel perspective towards understanding CRYPα's functions and regulation has been opened. In addition, it became apparent that this RPTP itself can be phosphorylated in vivo and preliminary experiments suggest the existence of a ~75 kDa putative substrate for its enzymatic activity. Altogether, these data contribute to the molecular characterisation of CRYPα and suggest potential ways to modulate its signalling function in pathological conditions.

الوصول الحر: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.271502Test