DOCSLIB.ORG

Synthesis of Recombinant Human Relaxin H2 in Bacteria and Its Effects on Differentiation of Carcinoembryonic Stem Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Synthesis of Recombinant Human Relaxin H2 in Bacteria and Its Effects on Differentiation of Carcinoembryonic Stem Cells Synthesis of recombinant human relaxin H2 in bacteria and its effects on differentiation of carcinoembryonic stem cells by Roman Stanisław Poterski A Thesis presented to The University of Guelph In partial fulfillment of requirements for the degree of Doctor of Philosophy In Biomedical Sciences Guelph, Ontario, Canada © Roman Stanisław Poterski, 2017 ABSTRACT SYMTHESIS OF RECOMBINANT HUMAN RELAXIN H2 IN BACTERIA AND ITS EFFECTS ON DIFFERENTIATION OF CARCINOEMBRYONIC STEM CELLS Roman Stanislaw Poterski Co-Advisors: University of Guelph, 2017 Doctor Alastair J.S. Summerlee Doctor Tarek Saleh Experiments described in this thesis were designed to develop a reliable method for the synthesis of the recombinant human hormone relaxin H2. Several methods were tested to produce highly pure, biologically active protein quickly, inexpensively and consistently from batch to batch. Relaxin H2 synthesized in bacteria was assessed by SDS-PAGE gel electrophoresis, Western Dot Blotting, Liquid Chromatography/Ultra High Definition Mass Spectrometry, and in vivo blood pressure response experiments using rats. The recombinant relaxin H2 described in this study was compared with commercially available relaxin H2 and was determined to be of equal quality, purity, and biological activity. In addition, to determine whether or not relaxin H2 induced differentiation of murine carcinoembryonic stem cells P19CL6 into cardiomyocytes was compared with a standard methodology using DMSO-induced differentiation. Relaxin H2 induced differentiation but the onset was delayed by four days compared with DMSO treatment. The endpoint of differentiation was determined as the start of spontaneous contractions within clusters of cells in culture. The results revealed that both commercially obtained and bacterially produced relaxin H2 equally caused a four-day delay in the initiation of contractions by cardiac myocytes compared with DMSO-treated controls. Moreover, cDNA microarray analysis of P19CL6 cells in three separate cultures, induced with DMSO (1%, as a control), and experimental treatments with serelaxin (16.7 nM), and bacterially-derived RLN H2 (16.7nM), respectively. There were no differences observed between cell treatments with the serelaxin- and bacterially-derived RLN H2 in the activation/deactivation patterns of 125 affected genes during the progression of the differentiation process. ACKNOWLEDGEMENTS I would like to thank all of the many people who have supported me during the course of my doctoral project. First and foremost, I thank Doctor Alastair J. S. Summerlee for his guidance and unreserved support. Despite being incredibly busy with his administrative, teaching, research and humanitarian endeavors, Alastair always found time to answer my questions and provided advice. There are not many professors of Alastair’s standing who would take on a part-time graduate student and would also support the candidate’s full-time workload. There were also absences from my graduate program due to the extenuating circumstances. Thank you, Alastair, for your continued support. I thank Dr. Tarek Saleh for agreeing to serve as my co-advisor and for his support. The members of my Advisory Committee, Drs. Coral Murrant and Brian Wilson were so important that without their support this project would not have been successful. Thank you. Members of the Biomedical Sciences Department of whom there are too many to name all, thank you. Dr. Jeff Thomason and the anatomy team, thank you for your support and encouragement. The DVM students with whom I have contact during their anatomy laboratory classes have been my unwavering supporters. I especially thank OVC Classes of 2013 and 2018 for cheering me on and electing me their Honorary Class President. iv My family, Barbara, Marta, Dave, Noah, and Seth were the true pillars and foundation on which I was able to build my doctoral aspiration. Thank you all with all my heart. v DECLARATION OF WORK PERFORMED I declare that the work described in this thesis has been completed by me with the following exceptions requiring the expertise and equipment not available locally: Chemical synthesis and sequencing of DNA’s were performed by University of Guelph Molecular Laboratory Services. Mass spectrometry analyses were done by at the Advanced Analysis Centre, Mass Spectrometry Laboratory, University of Guelph. Microarray hybridizations and partial statistical analyses were completed with the assistance of University Health Network Microarray Centre, Toronto. vi Table of Contents ACKNOWLEDGEMENTS............................................................................................................... iv DECLARATION OF WORK PERFORMED ................................................................................... vi LIST OF FIGURES .......................................................................................................................... x LIST OF TABLES ......................................................................................................................... xiii LIST OF ABBREVIATIONS .......................................................................................................... xv INTRODUCTION .............................................................................................................................. 1 OUTLINE OF THE THESIS ............................................................................................................. 5 CHAPTER 1............................................................................................................. 7 REVIEW OF THE LITERATURE ...................................................................................................................... 7 The discovery of relaxin................................................................................................................... 9 Tissue distribution of relaxin isoforms and their receptors ................................................. 10 BIOLOGICAL ACTIONS OF RLN ................................................................................................................ 14 Central effects of relaxin ................................................................................................................ 14 Relaxin, the heart, and circulation .............................................................................................. 16 Tissue remodeling ........................................................................................................................... 18 Relaxin and cancer .......................................................................................................................... 20 Relaxin and cell differentiation .................................................................................................... 21 GENERAL STRATEGIES FOR PRODUCTION OF RECOMBINANT PROTEINS ................................................ 23 Production by secretion................................................................................................................. 26 Production within the cell .............................................................................................................. 28 Fusion proteins ................................................................................................................................ 29 Protein tags ....................................................................................................................................... 30 Bacterial culturing systems .......................................................................................................... 30 Cell-free synthesis of proteins ..................................................................................................... 31 THE PRINCIPLES OF DESIGN AND CONSTRUCTION OF VECTORS FOR BACTERIAL EXPRESSION OF RECOMBINANT PROTEINS ......................................................................................................................... 35 The rationale of design and construction of eukaryotic expression vectors .................. 44 Expression of proteins in mammalian cells ............................................................................. 47 Expression of proteins in E. coli .................................................................................................. 47 HISTORICAL SYNOPSIS OF METHODS USED FOR PRODUCTION OF RELAXIN............................................ 48 RATIONALE .................................................................................................................................. 51 CHAPTER 2 ................................................................................................................................... 54 EXPRESSION OF HUMAN RLN H2 IN E. COLI USING A HEXA-HISTIDINE TAG AND MALTOSE-BINDING PROTEIN DUAL-AFFINITY FUSION SYSTEM ......................................... 54 INTRODUCTION .......................................................................................................................................... 54 RATIONALE ............................................................................................................................................... 61 MATERIALS AND METHODS ....................................................................................................................... 62 Methodology used to prepare bench-top RLN H2 ................................................................... 62 Plasmids and bacterial strains ....................................................................................................
Load more

Recommended publications
  • Anthropoid Primate–Specific Retroviral Element THE1B Controls Expression of CRH in Placenta and Alters Gestation Length
    SHORT REPORTS Anthropoid primate±specific retroviral element THE1B controls expression of CRH in placenta and alters gestation length 1 1 1 2 Caitlin E. Dunn-FletcherID *, Lisa M. Muglia , Mihaela Pavlicev , Gernot Wolf , Ming- An Sun2, Yueh-Chiang Hu3, Elizabeth Huffman1, Shivani Tumukuntala1, Katri Thiele1, Amrita Mukherjee1, Sandra Zoubovsky1, Xuzhe Zhang1, Kayleigh A. Swaggart1, Katherine Y. Bezold Lamm1, Helen Jones4, Todd S. Macfarlan2, Louis J. Muglia1* a1111111111 1 Division of Human Genetics, Center for Prevention of Preterm Birth, Perinatal Institute, Cincinnati a1111111111 Children's Hospital Medical Center, Department of Pediatrics, University of Cincinnati College of Medicine, a1111111111 Cincinnati, Ohio, United States of America, 2 The Eunice Kennedy Shriver National Institute of Child Health a1111111111 and Human Development, The National Institutes of Health, Bethesda, Maryland, United States of America, 3 Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Department of a1111111111 Pediatrics, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America, 4 Division of Pediatric Surgery, Cincinnati Children's Hospital Medical Center, Department of Surgery, University of Cincinnati College of Medicine, Cincinnati, Ohio, United States of America * [email protected] (CED); [email protected] (LJM) OPEN ACCESS Citation: Dunn-Fletcher CE, Muglia LM, Pavlicev M, Wolf G, Sun M-A, Hu Y-C, et al. (2018) Anthropoid Abstract primate±specific retroviral element THE1B controls expression of CRH in placenta and alters gestation Pregnancy and parturition are intricately regulated to ensure successful reproductive out- length. PLoS Biol 16(9): e2006337. https://doi.org/ comes. However, the factors that control gestational length in humans and other anthropoid 10.1371/journal.pbio.2006337 primates remain poorly defined.
    [Show full text]
  • Guidelines on Pulmonary Hypertension 2015 (TF08) - Task Force Members and Additional Contributors
    Guidelines on Pulmonary Hypertension 2015 (TF08) - Task Force Members and Additional Contributors For ESC Guidelines: The report below lists declarations of interest as reported to the ESC by the experts covering the period of the Guidelines production, from Task Force creation to publication. Expert Type of Relationship with Industry Beghetti Maurice A - Direct Personal payment: Speaker fees, Honoraria, Consultancy, Advisory Board fees, Investigator, Committee Member, etc. - Novartis : Pulmonary hypertension imatinib (2012) - Pfizer : Pulmonary hypertension sildenafil (2012-2013) - Bayer Schering Pharma : Pulmonary hypertension riociguat (2012-2013-2014-2015) - Eli Lilly : Pulmonary hypertension tadalafil (2012-2013-2014-2015) - Actelion : Pulmonary hypertension, Tracleer, Macitentan, Selexipag (2012-2013-2014-2015) - GlaxoSmithKline : pulmonary hypertension Ambrisentan (2012-2015) - Novartis : Pulmonary hypertension riociguat (2013) - GlaxoSmithKline : ambrisentan (2014) D - Research funding (departmental or institutional). - Actelion : no relation to a specific product (2012-2013) - Bayer Schering Pharma : no relation to a specific product (2014-2015) Galie Nazzareno A - Direct Personal payment: Speaker fees, Honoraria, Consultancy, Advisory Board fees, Investigator, Committee Member, etc. - Eli Lilly : pulmonary hypertension (2012-2013) - Novartis : pulmonary hypertension (2012-2013) - Pfizer : pulmonary hypertension (2012-2013) - Actelion : pulmonary hypertension (2012-2013) - GlaxoSmithKline : pulmonary hypertension (2012-2013)
    [Show full text]
  • Future Diagnostic & Therapeutic Targets in Cardiorenal Syndromes
    Future Diagnostic & Therapeutic Targets in Cardiorenal Syndromes (Biomarkers, advanced monitoring, advanced imaging, novel therapies) EDGAR V. LERMA, MD Clinical Professor of Medicine Secon of Nephrology UIC/ Advocate Christ Medical Center Oak Lawn, IL May 27, 2017 Disclosure of Interests • Honoraria: UpToDate, McGraw-Hill Publishing, Elsevier Publishing, Springer Publishing, Wolters-Kluwer Publishing, ACP Smart Medicine, Emedicine • Editorial Boards: American Journal of Kidney Diseases, ASN Kidney News, Clinical Journal of the American Society of Nephrology, Clinical Reviews in Bone and Mineral Metabolism, International Urology and Nephrology, Journal of Clinical Lipidology, Prescribers Letter, Renal and Urology News, Reviews in Endocrinology and Metabolic Disorders, Seminars in Dialysis • Speaker/ Advisory Board: Astute Medical, Mallinckrodt, Otsuka Pharmaceuticals, ZS Pharma KDIGO Controversies Conference on Heart Failure in CKD May 25-28, 2017 | Athens, Greece Disclosure of ABIM Service: Edgar V. Lerma, M.D. ▪ I am a current member of the ABIM Self-Assessment Committee. ▪ To protect the integrity of certification, ABIM enforces strict confidentiality and ownership of exam content. ▪ My participation in this CME activity is allowed under ABIM policy and is subject to the following: • As a member of an ABIM test committee, I agreed to keep exam information confidential, as it is owned exclusively by ABIM. • As is true for any ABIM candidate who has taken an exam for certification, I have signed the Pledge of Honesty in which I have agreed
    [Show full text]
  • The Expression of Human Endogenous Retroviruses Is Modulated by the Tat Protein of HIV‐1
    The Expression of Human Endogenous Retroviruses is modulated by the Tat protein of HIV‐1 by Marta Jeannette Gonzalez‐Hernandez A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Immunology) in The University of Michigan 2012 Doctoral Committee Professor David M. Markovitz, Chair Professor Gary Huffnagle Professor Michael J. Imperiale Associate Professor David J. Miller Assistant Professor Akira Ono Assistant Professor Christiane E. Wobus © Marta Jeannette Gonzalez‐Hernandez 2012 For my family and friends, the most fantastic teachers I have ever had. ii Acknowledgements First, and foremost, I would like to thank David Markovitz for his patience and his scientific and mentoring endeavor. My time in the laboratory has been an honor and a pleasure. Special thanks are also due to all the members of the Markovitz laboratory, past and present. It has been a privilege, and a lot of fun, to work near such excellent scientists and friends. You all have a special place in my heart. I would like to thank all the members of my thesis committee for all the valuable advice, help and jokes whenever needed. Our collaborators from the Bioinformatics Core, particularly James Cavalcoli, Fan Meng, Manhong Dai, Maureen Sartor and Gil Omenn gave generous support, technical expertise and scientific insight to a very important part of this project. Thank you. Thanks also go to Mariana Kaplan’s and Akira Ono’s laboratory for help with experimental designs and for being especially generous with time and reagents. iii Table of Contents Dedication ............................................................................................................................ ii Acknowledgements ............................................................................................................. iii List of Figures ...................................................................................................................
    [Show full text]
  • Classification Decisions Taken by the Harmonized System Committee from the 47Th to 60Th Sessions (2011
    CLASSIFICATION DECISIONS TAKEN BY THE HARMONIZED SYSTEM COMMITTEE FROM THE 47TH TO 60TH SESSIONS (2011 - 2018) WORLD CUSTOMS ORGANIZATION Rue du Marché 30 B-1210 Brussels Belgium November 2011 Copyright © 2011 World Customs Organization. All rights reserved. Requests and inquiries concerning translation, reproduction and adaptation rights should be addressed to [email protected]. D/2011/0448/25 The following list contains the classification decisions (other than those subject to a reservation) taken by the Harmonized System Committee ( 47th Session – March 2011) on specific products, together with their related Harmonized System code numbers and, in certain cases, the classification rationale. Advice Parties seeking to import or export merchandise covered by a decision are advised to verify the implementation of the decision by the importing or exporting country, as the case may be. HS codes Classification No Product description Classification considered rationale 1. Preparation, in the form of a powder, consisting of 92 % sugar, 6 % 2106.90 GRIs 1 and 6 black currant powder, anticaking agent, citric acid and black currant flavouring, put up for retail sale in 32-gram sachets, intended to be consumed as a beverage after mixing with hot water. 2. Vanutide cridificar (INN List 100). 3002.20 3. Certain INN products. Chapters 28, 29 (See “INN List 101” at the end of this publication.) and 30 4. Certain INN products. Chapters 13, 29 (See “INN List 102” at the end of this publication.) and 30 5. Certain INN products. Chapters 28, 29, (See “INN List 103” at the end of this publication.) 30, 35 and 39 6. Re-classification of INN products.
    [Show full text]
  • The Evolution of Heart Failure with Reduced Ejection Fraction Pharmacotherapy: What Do We Have and Where Are We Going?
    Pharmacology & Therapeutics 178 (2017) 67–82 Contents lists available at ScienceDirect Pharmacology & Therapeutics journal homepage: www.elsevier.com/locate/pharmthera Associate editor: M. Curtis The evolution of heart failure with reduced ejection fraction pharmacotherapy: What do we have and where are we going? Ahmed Selim, Ronald Zolty, Yiannis S. Chatzizisis ⁎ Division of Cardiovascular Medicine, University of Nebraska Medical Center, Omaha, NE, USA article info abstract Available online 21 March 2017 Cardiovascular diseases represent a leading cause of mortality and increased healthcare expenditure worldwide. Heart failure, which simply describes an inability of the heart to meet the body's needs, is the end point for many Keywords: other cardiovascular conditions. The last three decades have witnessed significant efforts aiming at the discovery Heart failure of treatments to improve the survival and quality of life of patients with heart failure; many were successful, Reduced ejection fraction while others failed. Given that most of the successes in treating heart failure were achieved in patients with re- Pharmacotherapy duced left ventricular ejection fraction (HFrEF), we constructed this review to look at the recent evolution of Novel drugs HFrEF pharmacotherapy. We also explore some of the ongoing clinical trials for new drugs, and investigate poten- tial treatment targets and pathways that might play a role in treating HFrEF in the future. © 2017 Elsevier Inc. All rights reserved. Contents 1. Introduction..............................................
    [Show full text]
  • Involvement of Cyclic Guanosine Monophosphate-Dependent Protein Kinase I in Renal Antifibrotic Effects of Serelaxin
    fphar-07-00195 July 9, 2016 Time: 13:1 # 1 View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Frontiers - Publisher Connector ORIGINAL RESEARCH published: 12 July 2016 doi: 10.3389/fphar.2016.00195 Involvement of Cyclic Guanosine Monophosphate-Dependent Protein Kinase I in Renal Antifibrotic Effects of Serelaxin Veronika Wetzl1,2†, Elisabeth Schinner1†, Frieder Kees1, Franz Hofmann3, Lothar Faerber1,2 and Jens Schlossmann1* 1 Department of Pharmacology and Toxicology, University of Regensburg, Regensburg, Germany, 2 Novartis Pharma GmbH, Nuremberg, Germany, 3 Institute of Pharmacology and Toxicology, Technical University of Munich, Munich, Germany Introduction: Kidney fibrosis has shown to be ameliorated through the involvement of cyclic guanosine monophosphate (cGMP) and its dependent protein kinase I (cGKI). Serelaxin, the recombinant form of human relaxin-II, increases cGMP levels and has Edited by: Enno Klussmann, shown beneficial effects on kidney function in acute heart failure patients. Antifibrotic Max Delbrüeck Center for Molecular properties of serelaxin are supposed to be mediated via relaxin family peptide receptor Medicine, Germany 1 and subsequently enhanced nitric oxide/ cGMP to inhibit transforming growth factor- Reviewed by: Friedrich Wilhelm Herberg, b (TGF-b) signaling. This study examines the involvement of cGKI in the antifibrotic University of Kassel, Germany signaling of serelaxin. Choel Kim, Baylor College of Medicine, USA Methods and Results: Kidney fibrosis was induced by unilateral ureteral obstruction in *Correspondence: wildtype (WT) and cGKI knock-out (KO) mice. After 7 days, renal antifibrotic effects Jens Schlossmann of serelaxin were assessed. Serelaxin treatment for 7 days significantly increased [email protected] regensburg.de cGMP in the kidney of WT and cGKI-KO.
    [Show full text]
  • Expression of Placental Regulatory Genes Is Associated with Fetal Growth
    City University of New York (CUNY) CUNY Academic Works Publications and Research Queens College 2017 Expression of Placental Regulatory Genes Is Associated with Fetal Growth Maya A. Deyssenroth Icahn School of Medicine at Mount Sinai Qian Li Icahn School of Medicine at Mount Sinai Marina Lacasaña Andalusian School of Public Health Yoko Nomura CUNY Queens College Carmen Marsit Emory University See next page for additional authors How does access to this work benefit ou?y Let us know! More information about this work at: https://academicworks.cuny.edu/qc_pubs/189 Discover additional works at: https://academicworks.cuny.edu This work is made publicly available by the City University of New York (CUNY). Contact: [email protected] Authors Maya A. Deyssenroth, Qian Li, Marina Lacasaña, Yoko Nomura, Carmen Marsit, and Jia Chen This article is available at CUNY Academic Works: https://academicworks.cuny.edu/qc_pubs/189 J. Perinat. Med. 2017; aop Maya A. Deyssenrotha, Qian Lia, Marina Lacasaña, Yoko Nomura, Carmen Marsit and Jia Chen* Expression of placental regulatory genes is associated with fetal growth DOI 10.1515/jpm-2017-0064 independent cohort (n = 306). Given that aberrant fetal Received February 25, 2017. Accepted May 31, 2017. growth may have long-lasting effects, our results suggest the potential utility of placental gene expression profiles as Abstract: The placenta is the principal organ regulating potential early markers of disease onset later in life. respiratory, nutritional, endocrine and metabolic functions on behalf of the developing fetus. Changes in gene expres- Keywords: Birth weight; gene expression, placenta. sion patterns of placenta-specific genes may influence fetal growth.
    [Show full text]
  • Jimmunol.1800856.Full.Pdf
    Healthy Donors Exhibit a CD4 T Cell Repertoire Specific to the Immunogenic Human Hormone H2-Relaxin before Injection This information is current as of September 26, 2021. Aurélien Azam, Yann Gallais, Sergio Mallart, Stephane Illiano, Olivier Duclos, Catherine Prades and Bernard Maillère J Immunol published online 17 May 2019 http://www.jimmunol.org/content/early/2019/05/14/jimmun Downloaded from ol.1800856 Supplementary http://www.jimmunol.org/content/suppl/2019/05/14/jimmunol.180085 http://www.jimmunol.org/ Material 6.DCSupplemental Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists by guest on September 26, 2021 • Fast Publication! 4 weeks from acceptance to publication *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2019 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Published May 17, 2019, doi:10.4049/jimmunol.1800856 The Journal of Immunology Healthy Donors Exhibit a CD4 T Cell Repertoire Specific to the Immunogenic Human Hormone H2-Relaxin before Injection Aure´lien Azam,*,† Yann Gallais,† Sergio Mallart,‡ Stephane Illiano,x Olivier Duclos,‡ Catherine Prades,* and Bernard Maille`re† H2-relaxin (RLN2) is a two-chain peptide hormone structurally related to insulin with a therapeutic potential in multiple indica- tions.
    [Show full text]
  • ATC/DDD Classification
    WHO Drug Information Vol. 27, No. 2, 2013 ATC/DDD Classification ATC/DDD Classification (Temporary) The following anatomical therapeutic codes (ATC), defined daily doses (DDD) and alterations were considered by the WHO International Working Group for Drug Statistics Methodology at its meeting in March 2013. Comments or objections to the decisions should be forwarded to the WHO Collaborating Centre for Drug Statistics Methodology at [email protected]. The new ATC codes, DDDs and alterations will then be considered final and be included in the January 2014 version of the ATC/DDD index. The inclusion of a substance in the lists does not imply any recommendation for use in medicine or pharmacy. New ATC 5th level codes: ATC level INN/Common name ATC code afamelanotide D02BB02 apremilast L04AA32 brimonidine D11AX21 calcium citrate A12AA13 carfilzomib L01XX45 colestilan V03AE06 delamanid G04AK06 dienogest and ethinylestradiol G03AA16 eliglustat A16AX10 empagliflozin A10BX12 encephalitis, Japanese, live attenuated J07BA03 formoterol and fluticasone R03AK11 insulin degludec A10AE06 insulin degludec and insulin aspart A10AD05 lamivudine, tenofovir disoproxil and efavirenz J05AR11 macitentan C02KX04 metformin and dapagliflozin A10BD15 nalmefene N07BB05 naloxegol A06AH03 nomegestrol and estrogen G03FB12 obinutuzumab L01XC15 ocriplasmin S01AX22 ospemifene G03XC05 Continued/ 130 WHO Drug Information Vol. 27, No. 2, 2013 ATC/DDD Classification ATC level INN/Common name ATC code pomalidomide L04AX06 serelaxin C01DX21 strontium ranelate and colecalciferol
    [Show full text]
  • Supplementary Table 1
    Supplementary Table 1. 492 genes are unique to 0 h post-heat timepoint. The name, p-value, fold change, location and family of each gene are indicated. Genes were filtered for an absolute value log2 ration 1.5 and a significance value of p ≤ 0.05. Symbol p-value Log Gene Name Location Family Ratio ABCA13 1.87E-02 3.292 ATP-binding cassette, sub-family unknown transporter A (ABC1), member 13 ABCB1 1.93E-02 −1.819 ATP-binding cassette, sub-family Plasma transporter B (MDR/TAP), member 1 Membrane ABCC3 2.83E-02 2.016 ATP-binding cassette, sub-family Plasma transporter C (CFTR/MRP), member 3 Membrane ABHD6 7.79E-03 −2.717 abhydrolase domain containing 6 Cytoplasm enzyme ACAT1 4.10E-02 3.009 acetyl-CoA acetyltransferase 1 Cytoplasm enzyme ACBD4 2.66E-03 1.722 acyl-CoA binding domain unknown other containing 4 ACSL5 1.86E-02 −2.876 acyl-CoA synthetase long-chain Cytoplasm enzyme family member 5 ADAM23 3.33E-02 −3.008 ADAM metallopeptidase domain Plasma peptidase 23 Membrane ADAM29 5.58E-03 3.463 ADAM metallopeptidase domain Plasma peptidase 29 Membrane ADAMTS17 2.67E-04 3.051 ADAM metallopeptidase with Extracellular other thrombospondin type 1 motif, 17 Space ADCYAP1R1 1.20E-02 1.848 adenylate cyclase activating Plasma G-protein polypeptide 1 (pituitary) receptor Membrane coupled type I receptor ADH6 (includes 4.02E-02 −1.845 alcohol dehydrogenase 6 (class Cytoplasm enzyme EG:130) V) AHSA2 1.54E-04 −1.6 AHA1, activator of heat shock unknown other 90kDa protein ATPase homolog 2 (yeast) AK5 3.32E-02 1.658 adenylate kinase 5 Cytoplasm kinase AK7
    [Show full text]
  • Synthesis, Conformation, and Activity of Human Insulin- Like Peptide 5 (INSL5) Mohammed Akhter Hossain,[A] Ross A
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by PubMed Central DOI: 10.1002/cbic.200800113 Synthesis, Conformation, and Activity of Human Insulin- Like Peptide 5 (INSL5) Mohammed Akhter Hossain,[a] Ross A. D. Bathgate,[a, b] Chze K. Kong,[a, b] Fazel Shabanpoor,[a, c] Suode Zhang,[a] Linda M. Haugaard-Jçnsson,[d] K. Johan Rosengren,[d] Geoffrey W. Tregear,[a, b] and John D. Wade*[a, c] Insulin-like peptide 5 (INSL5) was first identified through searches resistant to standard synthesis protocols and required highly opti- of the expressed sequence tags (EST) databases. Primary se- mized conditions for their acquisition. In particular, the use of a quence analysis showed it to be a prepropeptide that was pre- strong tertiary amidine, DBU, as Na-deprotection base was re- dicted to be processed in vivo to yield a two-chain sequence (A quired for the successful assembly of the B chain; this highlights and B) that contained the insulin-like disulfide cross-links. The the need to consider incomplete deprotection rather than acyla- high affinity interaction between INSL5 and the receptor RXFP4 tion as a cause of failed synthesis. Following sequential disulfide (GPCR142) coupled with their apparent coevolution and partially bond formation and chain combination, the resulting synthetic overlapping tissue expression patterns strongly suggest that INSL5, which was obtained in good overall yield, was shown to INSL5 is an endogenous ligand for RXFP4. Given that the primary possess a similar secondary structure to human relaxin-3 (H3 re- function of the INSL5–RXFP4 pair remains unknown, an effective laxin).
    [Show full text]