DOCSLIB.ORG

(12) Patent Application Publication (10) Pub. No.: US 2016/0051674 A1 LATOUR Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(12) Patent Application Publication (10) Pub. No.: US 2016/0051674 A1 LATOUR Et Al US 2016.0051674A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0051674 A1 LATOUR et al. (43) Pub. Date: Feb. 25, 2016 (54) METHODS AND PHARMACEUTICAL A6II 45/06 (2006.01) COMPOSITIONS (CTPS1 INHIBITORS, E.G. A613 L/655 (2006.01) NORLEUCINE) FOR INHIBITING T CELL A613 L/436 (2006.01) PROLIFERATION IN A SUBJECT IN NEED A63L/85 (2006.01) THEREOF A613 L/353 (2006.01) A613 L/404 (2006.01) (71) Applicants: INSERM (INSTITUT NATIONAL DE A63L/215 (2006.01) LASANTE ET DE LA RECHERCHE A613 L/405 (2006.01) MEDICALE), Faris (FR): A 6LX3/5377 (2006.01) FONDATION IMAGINE, Paris (FR): A613 L/52 (2006.01) UNIVERSITE PARIS DESCARTES, A638/3 (2006.01) Paris (FR), ASSISTANCE GOIN33/50 (2006.01) PUBLIQUE-HOPITAUX DE PARIS A613 L/42 (2006.01) (APHP), Paris (FR): THE (52) U.S. Cl. UNIVERSTY OF MANCHESTER, CPC ............. A61K 39/3955 (2013.01); A61K3I/42 Manchester (GB) (2013.01); A61 K3I/7068 (2013.01); A61 K 3 1/713 (2013.01); A61K 45/06 (2013.01); (72) Inventors: Sylvain LATOUR, Paris (FR); Alain A6 IK3I/655 (2013.01); A61 K3I/436 FISCHER, Paris (FR); Emmanuel (2013.01); A61 K3I/I85 (2013.01); A61 K MARTIN, Paris (FR); Peter 31/353 (2013.01); A61K 31/404 (2013.01); ARKWRIGHT, Manchester (GB) A6 IK3I/215 (2013.01); A61 K3I/4015 (2013.01); A61 K3I/5377 (2013.01); A61 K (21) Appl. No.: 14/781,353 3 1/52 (2013.01); A61K 38/13 (2013.01); G0IN (22) PCT Filed: Apr. 17, 2014 33/505 (2013.01); G0IN33/5023 (2013.01) (57) ABSTRACT (86). PCT No.: PCT/EP2014/057895 The present invention relates to methods and pharmaceutical S371 (c)(1), compositions for inhibiting lymphocyte proliferationina Sub (2) Date: Sep. 30, 2015 ject in need thereof. In particular, the invention relates to a CTP synthase 1 (CTPS1) inhibitor for use in a method for (30) Foreign Application Priority Data inhibiting lymphocyte proliferationin a subject in need thereof. The invention also relates to a method for screening Apr. 18, 2013 (EP) .................................. 13305504.6 a plurality of test substances useful for inhibiting lymphocyte proliferationin a Subject in need thereof comprising the steps Publication Classification consisting of i) testing each of the test Substances for its ability to inhibit CTPS1 activity or expression and ii) identi (51) Int. Cl. fying the test substance which inhibits CTPS1 activity or A 6LX39/395 (2006.01) expression thereby to identify a test substance useful for A6 IK3I/7068 (2006.01) inhibiting lymphocyte proliferationin a subject in need A6 IK3I/713 (2006.01) thereof. Patent Application Publication Feb. 25, 2016 Sheet 1 of 5 US 2016/0051674 A1 sh scramblethis shCTPS1 Sh CelTrace violet : O arti-CTPS1 anti-CTPS at-ACTiN Figure 1A Patent Application Publication Feb. 25, 2016 Sheet 2 of 5 US 2016/0051674 A1 5 1. P1.2 O, 5 fw CeTrace wiolet -- anti-CD3 gmi) Figure 1B Ct. P1.2 C a -EptyEct. wisco :9 9 g HHCTFS wer E H. g 50 - |P12 A. al-re 30 art-CTPS1 g 20 Unplus anti-CTPS2 2 3 4. s 7 8 Mun at ACTN Time (days) Figure 1C Patent Application Publication Feb. 25, 2016 Sheet 3 of 5 US 2016/0051674 A1 d UTP +cytidine GTP 9 ATP Ctrl. no stim. Oanti-CD3/CD28 anti-CD3/CD28 +rucleotide orrucleoside P2 CelTracgwilet - Figure 1D 6 Cir. P1.2 CCtrl+CTP Cir. -Cytidine P.12+CTP P1.2+Cytidine 4. O 2O stir. anti-CD3 Figure 1E Patent Application Publication Feb. 25, 2016 Sheet 4 of 5 US 2016/0051674 A1 - Deazauridre - guanosine - adenosine + 4 nucleosides no stim. anti-CD3/CD28 anti-CD3/CD28 +nucleoside CalTraia villet he Figure 1F H 9 r 12 120 Ctr. Ctrl+ Deaza. X 5 80 80 P12 5 b 4. 4. OAZ 's no stir. anti-CD3f28 Figure 1G Patent Application Publication Feb. 25, 2016 Sheet 5 of 5 US 2016/0051674 A1 h Hf - Y 300 - E 'y g 200 v. - O - Y W - Cir. C) Pat. Pat.-CTPS1 Figure 1H U.75 3 9. 3. 5 -- P1.2P2.1 |Empty Wector ? L HH s 25 O 5 O 5 20 25 30 Time (days) Figure 1 US 2016/005 1674 A1 Feb. 25, 2016 METHODS AND PHARMACEUTICAL osphate (CTP) is a precursor required for the metabolism of COMPOSITIONS (CTPS1 INHIBITORS, E.G. DNA, RNA and phospholipids". CTP originates from two NORLEUCINE) FOR INHIBITING T CELL Sources: a Salvage pathway and a de novo synthesis pathway PROLIFERATION IN A SUBJECT IN NEED that depends on two enzymes, the CTP synthase (or syn THEREOF thetase) 1 and 2 (CTPS1 and CTPS2), although their respec tive roles are not known 7. CTP synthase activity is a poten FIELD OF THE INVENTION tially important step for DNA synthesis in lymphocytes. 0001. The present invention relates to methods and phar Here, the inventors report the identification of a loss of func maceutical compositions for inhibiting lymphocyte prolifera tion mutation (rs145092287) in CTPS1 in humans causing a novel and life threatening immunodeficiency characterized tion in a subject in need thereof. by an impaired capacity of activated T and B cells to prolif BACKGROUND OF THE INVENTION erate. Proliferation in response to antigen receptor-mediated activation is defective in CTPS1-deficient subject T and B 0002 Lymphocyte proliferation is the normal component cells or in normal T cells knocked-down with shRNA for of the immune reaction toward an antigen (e.g. a pathogen CTPS1. In contrast, proximal and distal TCR signaling events antigen). However in certain circumstances lymphocyte pro and responses were only weakly affected by the absence of liferation appears deleterious. For example, organ transplan CTPS1. Normal T-cell proliferation was restored in CTPS1 tation elicits a complex series of immunologic processes that deficient cells by expressing wild-type CTPS1 or by addition are generally categorized as inflammation, immunity, tissue of exogenous CTP or its nucleoside precursor, cytidine. repair and structural reinforcement of damaged tissues. Typi CTPS1 expression was found to be low in resting T cells, but cally T cell proliferation leads to inflammation by the secre rapidly upregulated following TCR activation. These results tion of proinflammatory cytokines, e.g., interleukin-2 (IL-2) highlight a key and specific role of CTPS1 in the immune and IFN-g. Accordingly, the skilled man in the art has tried to system by its capacity to Sustain the proliferation of activated develop immunosuppressive agents. Immunosuppressive lymphocytes during the immune response. CTPS1 may there drugs fall into five groups: (i) regulators of gene expression; fore represent a therapeutic target of immunosuppressive (ii) alkylating agents; (iii) inhibitors of de novo purine Syn drugs that could specifically dampen lymphocyte activation. thesis; (iv) inhibitors of denovo pyrimidine synthesis; and (v) 0005 Accordingly a first aspect of the invention relates to inhibitors of kinases and phosphatases. For example, gluco a method for reducing or inhibiting lymphocyte proliferation corticoids exert immunosuppressive and anti-inflammatory in a subject in need thereof comprising administering the activity mainly by inhibiting the expression of the genes for subject with a therapeutically effective amount of at least one IL-2 and other mediators. Methotrexate and its polyglutamate derivatives Suppress inflammatory responses through release CTP synthase 1 (CTPS1) inhibitor. of adenosine. Mycophenolic acid and mizoribine inhibit 0006. In some embodiments, the method of the present inosine monophosphate dehydrogenase. Mycophenolic acid invention is suitable for inhibiting or reducing T cell prolif induces apoptosis of activated T-lymphocytes. Cyclosporine eration. and FK-506/Tacrolimus inhibit the phosphatase activity of 0007. In some embodiments, the method of the present calcineurin. Rapamycin inhibits signal transduction from the invention is suitable for inhibiting or reducing B cell prolif IL-2, epidermal growth factor and other cytokine receptors. eration. Immunosuppressive and anti-inflammatory compounds in 0008. In some embodiments, the subject is a transplanted development include inhibitors of p38 kinase and of the type Subject. Typically the Subject may have been transplanted IV isoform of cyclic AMP phosphodiesterase, which is with a graft selected from the group consisting of heart, expressed in T cells. However, immunosuppressive agents are kidney, lung, liver, pancreas, pancreatic islets, brain tissue, associated with toxicity due to their nonspecific immunosup stomach, large intestine, Small intestine, cornea, skin, tra pressive effects. Reducing immunosuppression can prevent chea, bone, bone marrow, muscle, or bladder. The method of side effects related to over-immunosuppression. However, the invention is indeed particularly suitable for preventing or since the intrinsic immunosuppressive requirements for each Suppressing an immune response associated with rejection of donor recipient pair are unknown, immunosuppressive mini a donor tissue, cell, graft, or organ transplant by a recipient mization carries a potential risk of under-immunosuppression subject. Graft-related diseases or disorders include graft ver and consequent acute rejection, premature graft loss and sus host disease (GVDH), such as associated with bone mar death. A promising future application of immunosuppressive row transplantation, and immune disorders resulting from or drugs is to search for agents that inhibit lymphocyte prolif associated with rejection of organ, tissue, or cell graft trans eration by novel mechanisms, as the currently used agents, plantation (e.g., tissue or cell allografts or Xenografts), which all possess non-specific broad immunosuppressive including, e.g., grafts of skin, muscle, neurons, islets, organs, effects. parenchymal cells of the liver, etc. With regard to a donor tissue, cell, graft or Solid organ transplant in a recipient Sub SUMMARY OF THE INVENTION ject, it is believed that CTPS1 inhibitor according to the invention may be effective in preventing acute rejection of 0003. The present invention relates to methods and phar Such transplant in the recipient and/or for long-term mainte maceutical compositions for inhibiting lymphocyte prolifera nance therapy to prevent rejection of Such transplant in the tion in a subject in need thereof.
Load more

Recommended publications
  • Noelia Díaz Blanco
    Effects of environmental factors on the gonadal transcriptome of European sea bass (Dicentrarchus labrax), juvenile growth and sex ratios Noelia Díaz Blanco Ph.D. thesis 2014 Submitted in partial fulfillment of the requirements for the Ph.D. degree from the Universitat Pompeu Fabra (UPF). This work has been carried out at the Group of Biology of Reproduction (GBR), at the Department of Renewable Marine Resources of the Institute of Marine Sciences (ICM-CSIC). Thesis supervisor: Dr. Francesc Piferrer Professor d’Investigació Institut de Ciències del Mar (ICM-CSIC) i ii A mis padres A Xavi iii iv Acknowledgements This thesis has been made possible by the support of many people who in one way or another, many times unknowingly, gave me the strength to overcome this "long and winding road". First of all, I would like to thank my supervisor, Dr. Francesc Piferrer, for his patience, guidance and wise advice throughout all this Ph.D. experience. But above all, for the trust he placed on me almost seven years ago when he offered me the opportunity to be part of his team. Thanks also for teaching me how to question always everything, for sharing with me your enthusiasm for science and for giving me the opportunity of learning from you by participating in many projects, collaborations and scientific meetings. I am also thankful to my colleagues (former and present Group of Biology of Reproduction members) for your support and encouragement throughout this journey. To the “exGBRs”, thanks for helping me with my first steps into this world. Working as an undergrad with you Dr.
    [Show full text]
  • Structures, Functions, and Mechanisms of Filament Forming Enzymes: a Renaissance of Enzyme Filamentation
    Structures, Functions, and Mechanisms of Filament Forming Enzymes: A Renaissance of Enzyme Filamentation A Review By Chad K. Park & Nancy C. Horton Department of Molecular and Cellular Biology University of Arizona Tucson, AZ 85721 N. C. Horton ([email protected], ORCID: 0000-0003-2710-8284) C. K. Park ([email protected], ORCID: 0000-0003-1089-9091) Keywords: Enzyme, Regulation, DNA binding, Nuclease, Run-On Oligomerization, self-association 1 Abstract Filament formation by non-cytoskeletal enzymes has been known for decades, yet only relatively recently has its wide-spread role in enzyme regulation and biology come to be appreciated. This comprehensive review summarizes what is known for each enzyme confirmed to form filamentous structures in vitro, and for the many that are known only to form large self-assemblies within cells. For some enzymes, studies describing both the in vitro filamentous structures and cellular self-assembly formation are also known and described. Special attention is paid to the detailed structures of each type of enzyme filament, as well as the roles the structures play in enzyme regulation and in biology. Where it is known or hypothesized, the advantages conferred by enzyme filamentation are reviewed. Finally, the similarities, differences, and comparison to the SgrAI system are also highlighted. 2 Contents INTRODUCTION…………………………………………………………..4 STRUCTURALLY CHARACTERIZED ENZYME FILAMENTS…….5 Acetyl CoA Carboxylase (ACC)……………………………………………………………………5 Phosphofructokinase (PFK)……………………………………………………………………….6
    [Show full text]
  • A Path Towards SARS-Cov-2 Attenuation: Metabolic Pressure on CTP Synthesis Rules the Virus Evolution
    bioRxiv preprint doi: https://doi.org/10.1101/2020.06.20.162933; this version posted June 21, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-ND 4.0 International license. 1 A path towards SARS-CoV-2 attenuation: metabolic pressure on CTP synthesis rules the virus evolution Zhihua Ou1,2, Christos Ouzounis3, Daxi Wang1,2, Wanying Sun1,2,4, Junhua Li1,2, Weijun Chen2,5*, Philippe Marlière6, Antoine Danchin7,8* 1. BGI-Shenzhen, Shenzhen 518083, China. 2. Shenzhen Key Laboratory of Unknown Pathogen Identification, BGI-Shenzhen, Shenzhen 518083, China. 3. Biological Computation and Process Laboratory, Centre for Research and Technology Hellas, Chemical Process and Energy Resources Institute, Thessalonica 57001, Greece 4. BGI Education Center, University of Chinese Academy of Sciences, Shenzhen, 518083, China. 5. BGI PathoGenesis Pharmaceutical Technology, BGI-Shenzhen, Shenzhen, China. 6. TESSSI, The European Syndicate of Synthetic Scientists and Industrialists, 81 rue Réaumur, 75002, Paris, France 7. Kodikos Labs, Institut Cochin, 24, rue du Faubourg Saint-Jacques Paris 75014, France. 8. School of Biomedical Sciences, Li KaShing Faculty of Medicine, Hong Kong University, 21 Sassoon Road, Pokfulam, Hong Kong. * To whom correspondence should be addressed Tel: +331 4441 2551; Fax: +331 4441 2559 E-mail: [email protected] Correspondence may also be addressed to [email protected] Keywords ABCE1; cytoophidia; innate immunity; Maxwell’s demon; Nsp1; phosphoribosyltransferase; queuine bioRxiv preprint doi: https://doi.org/10.1101/2020.06.20.162933; this version posted June 21, 2020.
    [Show full text]
  • Ehrlichia Chaffeensis
    RESEARCH ARTICLE Ehrlichia chaffeensis TRP47 enters the nucleus via a MYND-binding domain-dependent mechanism and predominantly binds enhancers of host genes associated with signal transduction, cytoskeletal organization, and immune response a1111111111 1 1 2 1 1 a1111111111 Clayton E. KiblerID , Sarah L. Milligan , Tierra R. Farris , Bing Zhu , Shubhajit Mitra , Jere a1111111111 W. McBride1,2,3,4,5* a1111111111 a1111111111 1 Department of Pathology, University of Texas Medical Branch, Galveston, Texas, United States of America, 2 Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, United States of America, 3 Center for Biodefense and Emerging Infectious Diseases, University of Texas Medical Branch, Galveston, Texas, United States of America, 4 Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas, United States of America, 5 Institute for Human Infections and Immunity, University of Texas Medical Branch, Galveston, Texas, United States of OPEN ACCESS America Citation: Kibler CE, Milligan SL, Farris TR, Zhu B, * [email protected] Mitra S, McBride JW (2018) Ehrlichia chaffeensis TRP47 enters the nucleus via a MYND-binding domain-dependent mechanism and predominantly binds enhancers of host genes associated with Abstract signal transduction, cytoskeletal organization, and immune response. PLoS ONE 13(11): e0205983. Ehrlichia chaffeensis is an obligately intracellular bacterium that establishes infection in https://doi.org/10.1371/journal.pone.0205983 mononuclear phagocytes through largely undefined reprogramming strategies including Editor: Gary M. Winslow, State University of New modulation of host gene transcription. In this study, we demonstrate that the E. chaffeensis York Upstate Medical University, UNITED STATES effector TRP47 enters the host cell nucleus and binds regulatory regions of host genes rele- Received: April 25, 2018 vant to infection.
    [Show full text]
  • 190323111.Pdf
    Characterization of polymers of nucleotide biosynthetic enzymes By Sajitha Anthony April, 2017 A dissertation presented to the faculty of Drexel University College of Medicine in partial fulfillment for the requirements for the degree of Doctor of Philosophy in Molecular and Cellular Biology and Genetics i ii ACKNOWLEDGEMENTS First and foremost, I would like to thank my mentor Dr. Jeffrey Peterson for all of his support and guidance throughout the five years that I have been in his lab. He has truly inspired me with his tremendous enthusiasm for science and his constant encouragement. He has changed, for the better, the way I approach and see science. I could not have asked for a better mentor. Secondly, I would like to thank my committee members for all of their great ideas and their never-ending support. I thank them for answering all of my questions so patiently and always listening to what I had to say. After every meeting I’ve had with them, I always felt encouraged and confident. I would also like to thank my collaborators over at the University of Washington—Justin Kollman, Anika Burrell, and Matthew Johnson. It has been such a pleasure working with them. The journey of discovery that we took together has been very exciting. This was my first collaboration, and they have made it such a memorable and positive experience. iii I would like to thank my lab members, in particular Alex for being an awesome cubemate, colleague, and friend. He has been the sounding board for so many of my project’s ideas over the years and has given such knowledgeable input.
    [Show full text]
  • The Emergence of Pattern Discovery Techniques in Computational Biology
    Metabolic Engineering, 2(3):159-177, July 2000 The Emergence Of Pattern Discovery Techniques In Computational Biology Isidore Rigoutsos1 Aris Floratos Laxmi Parida Yuan Gao Daniel Platt Bioinformatics and Pattern Discovery Group IBM TJ Watson Research Center Yorktown Heights, NY 10598. October 17, 1999 / Revised: March 24 2000 Abstract In the last few years, pattern discovery has been emerging as a generic tool of choice for tackling problems from the computational biology domain. In this presentation, and after defining the problem in its generality, we review some of the algorithms that have appeared in the literature, and describe several applications of pattern discovery to problems from computational biology. 1. Introduction Recent years have witnessed an emergence of pattern discovery methodologies for solving numerous tasks which arise in computational biology. Known also as ‘data mining’ approaches, they represent a novel approach for extracting useful information from databases containing various types of biological information. Initially, dynamic programming techniques were applied to the analysis of biological sequences and to the determination of sequence similarity between a query sequence and one or more biological sequences (DNA, proteins, and fragments) from a collection. Subsequent studies of such sequence similarity revealed conserved functional and structural signals, thus making the argument for the usefulness of such approaches. Research effort spanning almost two decades gave rise to a number of useful algorithms and an abundance of interesting scientific results [58,77,63,2]. Almost in parallel, researchers began looking into other approaches in an effort to develop concise consensus sequences that captured and represented regions of similarity across several sequences presumed to be related.
    [Show full text]
  • Dual Antimicrobial and Antiproliferative Activity of Tcpask Peptide Derived from a Tribolium Castaneum Insect Defensin
    microorganisms Article Dual Antimicrobial and Antiproliferative Activity of TcPaSK Peptide Derived from a Tribolium castaneum Insect Defensin Aida Robles-Fort 1, Inmaculada García-Robles 1 , Wasundara Fernando 2, David W. Hoskin 2,3,4, Carolina Rausell 1,† and María Dolores Real 1,*,† 1 Department of Genetics, University of Valencia, Burjassot, 46100 Valencia, Spain; [email protected] (A.R.-F.); [email protected] (I.G.-R.); [email protected] (C.R.) 2 Department of Pathology, Faculty of Medicine, Dalhousie University, Halifax, NS B3H 4R2, Canada; [email protected] (W.F.); [email protected] (D.W.H.) 3 Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University, Halifax, NS B3H 4R2, Canada 4 Department of Surgery, Faculty of Medicine, Dalhousie University, Halifax, NS B3H 4R2, Canada * Correspondence: [email protected]; Tel.: +34-96-354-3397 † These authors contributed equally to this work. Abstract: Antimicrobial peptides (AMPs) found in the innate immune system of a wide range of organisms might prove useful to fight infections, due to the reported slower development of resistance to AMPs. Increasing the cationicity and keeping moderate hydrophobicity of the AMPs have been described to improve antimicrobial activity. We previously found a peptide derived from the Tribolium castaneum insect defensin 3, exhibiting antrimicrobial activity against several human pathogens. Here, we analyzed the effect against Staphyloccocus aureus of an extended peptide Citation: Robles-Fort, A.; (TcPaSK) containing two additional amino acids, lysine and asparagine, flanking the former peptide García-Robles, I.; Fernando, W.; fragment in the original insect defensin 3 protein. TcPaSK peptide displayed higher antimicrobial Hoskin, D.W.; Rausell, C.; Real, M.D.
    [Show full text]
  • Investigation of New Candidate Genes in a Cohort of Patients with Familial Congenital Hypopituitarism And
    Investigation of new candidate genes in a cohort of patients with familial congenital hypopituitarism and associated disorders Louise Cheryl Gregory UCL Doctor of Philosophy Thesis 1 Declaration I, Louise Cheryl Gregory confirm that the work presented in this thesis is my own. Where information has been derived from other sources, I confirm that this has been indicated in the thesis. Publication arising from this thesis: Gregory, L.C., Humayun, K.N., Turton, J.P., McCabe, M.J., Rhodes, S.J., Dattani, M.T. 2015. Novel Lethal Form of Congenital Hypopituitarism Associated With the First Recessive LHX4 Mutation. J Clin Endocrinol Metab. 100(6): 2158-64. 2 Abstract Congenital hypopituitarism is a complex variable genetic disorder that is known to be caused by multiple mutated genes, both in isolation and in variably penetrant cases of digenic inheritance. In only <10% of cases, a mutation in a known causative gene has been identified in the patient, leaving the vast majority of patients yet to have a genetic mutation detected that is responsible for the pathogenicity and that has functional significance to their condition. This study investigates novel genes and pathways involved in hypothalamo-pituitary development. Our large cohort of consanguineous and non-consanguineous pedigrees with hypothalamo-pituitary disease are routinely screened for variants in the known causative genes. In pedigrees where there are no variants in these particular genes, exome sequencing in collaboration with GOSgene is carried out to uncover novel genes and regions of interest that are abnormal in the individual. Upon the identification of any novel variant in known or novel genes, functional assays are conducted to further show the significance of the change.
    [Show full text]
  • Coupled Structural Transitions Enable Highly Cooperative Regulation of 2 Human CTPS2 Filaments 3 4 Eric M
    bioRxiv preprint doi: https://doi.org/10.1101/770594; this version posted September 16, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Coupled structural transitions enable highly cooperative regulation of 2 human CTPS2 filaments 3 4 Eric M. Lynch1 & Justin M. Kollman1* 5 1Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA 6 *Corresponding author 7 8 Many enzymes assemble into defined oligomers, providing a mechanism for 9 cooperatively regulating enzyme activity. Recent studies in tissues, cells, and in vitro 10 have described a mode of regulation in which enzyme activity is modulated by 11 polymerization into large-scale filaments1-5. Enzyme polymerization is often driven by 12 binding to substrates, products, or allosteric regulators, and tunes enzyme activity by 13 locking the enzyme in high or low activity states1-5. Here, we describe a unique, 14 ultrasensitive form of polymerization-based regulation employed by human CTP 15 synthase 2 (CTPS2). High-resolution cryoEM structures of active and inhibited CTPS2 16 filaments reveal the molecular basis of this regulation. Rather than selectively stabilizing 17 a single conformational state, CTPS2 filaments dynamically switch between active and 18 inactive filament forms in response to changes in substrate and product levels. Linking 19 the conformational state of many CTPS2 subunits in a filament results in highly 20 cooperative regulation, greatly exceeding the limits of cooperativity for the CTPS2 21 tetramer alone. The structures also reveal a link between conformational state and 22 control of ammonia channeling between the enzyme’s two active sites.
    [Show full text]
  • Genetic Variation and Functional Analysis of the Cardiomedin Gene
    TECHNISCHE UNIVERSITÄT MÜNCHEN LEHRSTUHL FÜR EXPERIMENTELLE GENETIK Genetic Variation and Functional Analysis of the Cardiomedin Gene Zasie Susanne Schäfer Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzende: Univ.-Prof. A. Schnieke, Ph.D. Prüfer der Dissertation: 1. apl. Prof. Dr. J. Adamski 2. Univ.-Prof. Dr. Dr. H.-R. Fries 3. Univ.-Prof. Dr. Th. Meitinger Die Dissertation wurde am. 31.05.2011 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 02.04.2012 angenommen. Table of Contents Table of contents Abbreviations ........................................................................................................................ 7 1. Summary ..........................................................................................................................10 Zusammenfassung ...............................................................................................................11 2. Introduction ......................................................................................................................12 2.1 Genome-wide association studies (GWAS) and post-GWAS functional genomics ......12 2.2 Genetic influences on cardiac repolarization and sudden cardiac death syndrome in GWAS and the chromosome
    [Show full text]
  • Universidad Autónoma De Madrid
    Universidad Autónoma de Madrid Departamento de Bioquímica Identification of the substrates of the protease MT1-MMP in TNFα- stimulated endothelial cells by quantitative proteomics. Analysis of their potential use as biomarkers in inflammatory bowel disease. Agnieszka A. Koziol Tesis doctoral Madrid, 2013 2 Departamento de Bioquímica Facultad de Medicina Universidad Autónoma de Madrid Identification of the substrates of the protease MT1-MMP in TNFα- stimulated endothelial cells by quantitative proteomics. Analysis of their potential use as biomarkers in inflammatory bowel disease. Memoria presentada por Agnieszka A. Koziol licenciada en Ciencias Biológicas para optar al grado de Doctor. Directora: Dra Alicia García Arroyo Centro Nacional de Investigaciones Cardiovasculares (CNIC) Madrid, 2013 3 4 ACKNOWLEDGEMENTS - ACKNOWLEDGEMENTS - First and foremost I would like to express my sincere gratitude to my supervisor Dr. Alicia García Arroyo for giving me the great opportunity to study under her direction. Her encouragement, guidance and support from the beginning to the end, enabled me to develop and understand the subject. Her feedback to this manuscript, critical reading and corrections was inappreciable to finish this dissertation. Next, I would like to express my gratitude to those who helped me substantially along the way. To all members of MMPs’ lab, for their interest in the project, teaching me during al these years and valuable contribution at the seminary discussion. Without your help it would have not been possible to do some of those experiments. Thank you all for creating a nice atmosphere at work and for being so good friends in private. I would like to also thank all of the collaborators and technical units for their support and professionalism.
    [Show full text]
  • Regulation of Mammalian Nucleotide Metabolism and Biosynthesis Andrew N
    University of Kentucky UKnowledge Toxicology and Cancer Biology Faculty Toxicology and Cancer Biology Publications 2-27-2015 Regulation of Mammalian Nucleotide Metabolism and Biosynthesis Andrew N. Lane University of Kentucky, [email protected] Teresa W-M Fan University of Kentucky, [email protected] Click here to let us know how access to this document benefits oy u. Follow this and additional works at: https://uknowledge.uky.edu/toxicology_facpub Part of the Medical Toxicology Commons Repository Citation Lane, Andrew N. and Fan, Teresa W-M, "Regulation of Mammalian Nucleotide Metabolism and Biosynthesis" (2015). Toxicology and Cancer Biology Faculty Publications. 34. https://uknowledge.uky.edu/toxicology_facpub/34 This Article is brought to you for free and open access by the Toxicology and Cancer Biology at UKnowledge. It has been accepted for inclusion in Toxicology and Cancer Biology Faculty Publications by an authorized administrator of UKnowledge. For more information, please contact [email protected]. Regulation of Mammalian Nucleotide Metabolism and Biosynthesis Notes/Citation Information Published in Nucleic Acids Research, v. 43, issue 4, p. 2466-2485. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. Digital Object Identifier (DOI) http://dx.doi.org/10.1093/nar/gkv047 This article is available at UKnowledge: https://uknowledge.uky.edu/toxicology_facpub/34 2466–2485 Nucleic Acids Research, 2015, Vol.
    [Show full text]