DOCSLIB.ORG

Investigation of the Zinc Binding Region of Prothymosin-Alpha: a Spectroscopic and Thermodynamic Approach to Study Metal Binding in Intrinsically Disordered Proteins

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Investigation of the Zinc Binding Region of Prothymosin-Alpha: a Spectroscopic and Thermodynamic Approach to Study Metal Binding in Intrinsically Disordered Proteins Investigation of the Zinc Binding Region of Prothymosin-alpha: A Spectroscopic and Thermodynamic Approach to Study Metal Binding in Intrinsically Disordered Proteins Abstract The goal of this study is to provide a deeper insight into how metals bind intrinsically disordered proteins (IDPs) by taking zinc binding to prothymosin-alpha as a case study. The involvement of metals in several diseases caused by (IDPs) has been noted long ago (Cu binding of α-synuclein in Parkinsons disease, prion aggregation due to Cu in madcow disease. etc.). However, there is still a lack of understanding of how metal binds IDPs. A deeper insight into metal binding by IDPs is essential for a full understanding of a proteome and to pave a way for drug development. Prothymosin-alpha (Protα) is an acidic, natively unfolded, and highly conserved protein located mostly in the nucleus of eukaryotic cells. It consists of 110 amino acids out of which 53 residues are aspartic (D) and glutamic (E) acids thus it is highly acidic. The exact biological role of the protein is unknown. However, it has been shown to be involved in cell proliferation, chromatin remodeling, antiapoptotic activity etc. One of the most interesting characteristics of Protα is its ability to bind metal cations like Ca2+, Mg2+, Mn2+, Zn2+, Al2+. In the background of all these cations, Protα selectively interacts with Zn2+ and undergoes a structural change plus the interactions of the protein with its binding partners are enhanced in the presence of Zn2+. These features underline the importance of Zn2+ binding in Protα. Very little information is available about Zn2+ binding to Protα. A recent NMR study has shown that the 48-110 segment of Protα is the Zn2+ binding region of the protein. Although the 48-110 region is shown to be the Zn2+ binding region, the central 50-89 segment, where most of the Zn2+ binding residues are located in the protein, was used for all the studies here. To aid in the synthesis, purification, and characterization of highly negatively charged sequences like Protα(50-89)N50W new amino acid derivatives, 4-pyridylmethyl protected glutamic and aspartic acids were developed and their application was tested. Subsequently a circular dichroism (CD) spectroscopic study on Protα(50-89)N50W revealed that the peptide, like the full length protein, undergoes a structural change in the presence of Zn2+ and both the full protein and the peptide were saturated at 75 eq of Zn2+ indicating that the majority of Zn added binds in this region. The above study had also shown that selective binding of Zn2+ by Protα(50-89)N50W is due to sequence specificity and not solely because of electrostatic interactions between Protα(50- 89)N50W and Zn2+. Further the CD and differential scanning calorimetry (DSC) studies have shown that Protα(50-89)N50W undergoes a structural change similar to an alpha helix when heated from 20-80 oC. To set the stage for determining the specific glutamic and aspartic residues in Prot that act as ligands for Zn2+, a proof of principle for spin inversion recovery experiments has been provided using Zn2+ binding to EDTA as a model. This idea can further be extended to Protα(50- 89)N50W to understand if Zn2+ binds to a specific set of residues or if several degenerate binding sites exist. Investigation of the Zinc Binding Region of Prothymosin-alpha: A Spectroscopic and Thermodynamic Approach to Study Metal Binding in Intrinsically Disordered Proteins By SriRamya Garapati A Dissertation Submitted to the Faculty of East Carolina University In Partial Fulfillment of the Requirements For the Degree of Doctor of Philosophy In the Department of Chemistry East Carolina University, North Carolina 2014 ©SriRamya Garapati, 2014 Investigation of the Zinc Binding Region of Prothymosin-alpha: A Spectroscopic and Thermodynamic Approach to Study Metal Binding in Intrinsically Disordered Proteins By SriRamya Garapati APPROVED BY: DIRECTOR OF DISSERTATION: ______________________________________________________________________________ Colin Burns, Ph.D. COMMITTEE MEMBER: ______________________________________________________________________________ William Allen, Ph.D. COMMITTEE MEMBER: ______________________________________________________________________________ Art Rodriguez, Ph.D. COMMITTEE MEMBER: ______________________________________________________________________________ Anthony Kennedy, Ph.D. COMMITTEE MEMBER: ______________________________________________________________________________ Cindy Putnam Evans, Ph.D. DIRECTOR OF INTERDISCIPLINARY PROGRAM IN BIOLOGICAL SCIENCES: ______________________________________________________________________________ Terry West, Ph.D. DEAN OF THE GRADUATE SCHOOL: ______________________________________________________________________________ Paul Gemperline, Ph.D. DEDICATED To My beloved parents and husband Acknowledgements I heart fully thank my advisor, Dr. Colin Burns, without whom I could not have made through graduate school. His encouragement, guidance and support helped me in every way to develop an understanding of the subject. His patience and simplicity has always surprised me. I am very grateful for all his help in this journey I would like to extend my gratitude to Dr. Art Rodriguez and Dr. Anthony Kennedy who added their valuable suggestions and time to my research. I also thank my fellow graduate students for all the support they have offered throughout this journey. A special thanks to my committee members and Department of Chemistry for their assistance and time. Lastly, I offer my regards to all of those who supported me in any respect during the completion of the project. Table of Contents Dedication ......................................................................................................................................... ii Acknowledgements .......................................................................................................................... iii List of Figures .................................................................................................................................. vi List of Tables ................................................................................................................................. xiii List of Sequences ........................................................................................................................... xiv List of Schemes ............................................................................................................................... xv List of Abbreviation and Symbols ................................................................................................. xvi Chapter 1: Introduction 1.1 Natively Unfolded Proteins ........................................................................................................ 1 1.2 General Introduction About Prothymosin-α ............................................................................. 13 1.3 Biological Role of Prothymosin-α ........................................................................................... 15 1.4 Metal Binding Characteristics of Prothymosin-α .................................................................... 18 1.5 Research Goal and Overview of Research Strategy ................................................................ 26 Chapter 2: The 4-Pyridylmethyl Ester as a Protecting Group for Aspartic and Glutamic Acid: Flipping Peptide Charge States for Characterization by Positive Mode ESI-MS 2.1 Introduction .............................................................................................................................. 34 2.2 Research Strategy..................................................................................................................... 41 2.3 Synthesis, Purification and Characterization of 4-Pyridylmethyl Ester of Aspartic and Glutamic Acid ................................................................................................................................ 45 2.4 Application of 4-Pyridylmethyl Esters of Aspartic and Glutamic Acid in the Synthesis of Model Peptides................................................................................................................................ 50 2.5 Conclusions .............................................................................................................................. 57 Chapter 3: Zn and Temperature Induced Structural Changes in Protα(50-89)N50W Studied Using Circular Dichroism (CD) Spectroscopy 3.1 Introduction to CD Spectroscopy............................................................................................. 58 3.2 Research Strategy..................................................................................................................... 71 3.3 CD Measurements .................................................................................................................... 75 Chapter 4: Zn and Temperature Induced Structural Changes in Protα(50-89)N50W Studied Using Differential Scanning Calorimetry (DSC) 4.1 Introduction to DSC ................................................................................................................. 89 4.2 Research Strategy ..................................................................................................................... 99 4.3 DSC Measurements ............................................................................................................... 101 Chapter 5: Spin-Lattice Relaxation Measurements
Load more

Recommended publications
  • A Global Review on Short Peptides: Frontiers and Perspectives †
    molecules Review A Global Review on Short Peptides: Frontiers and Perspectives † Vasso Apostolopoulos 1 , Joanna Bojarska 2,* , Tsun-Thai Chai 3 , Sherif Elnagdy 4 , Krzysztof Kaczmarek 5 , John Matsoukas 1,6,7, Roger New 8,9, Keykavous Parang 10 , Octavio Paredes Lopez 11 , Hamideh Parhiz 12, Conrad O. Perera 13, Monica Pickholz 14,15, Milan Remko 16, Michele Saviano 17, Mariusz Skwarczynski 18, Yefeng Tang 19, Wojciech M. Wolf 2,*, Taku Yoshiya 20 , Janusz Zabrocki 5, Piotr Zielenkiewicz 21,22 , Maha AlKhazindar 4 , Vanessa Barriga 1, Konstantinos Kelaidonis 6, Elham Mousavinezhad Sarasia 9 and Istvan Toth 18,23,24 1 Institute for Health and Sport, Victoria University, Melbourne, VIC 3030, Australia; [email protected] (V.A.); [email protected] (J.M.); [email protected] (V.B.) 2 Institute of General and Ecological Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego˙ 116, 90-924 Lodz, Poland 3 Department of Chemical Science, Faculty of Science, Universiti Tunku Abdul Rahman, Kampar 31900, Malaysia; [email protected] 4 Botany and Microbiology Department, Faculty of Science, Cairo University, Gamaa St., Giza 12613, Egypt; [email protected] (S.E.); [email protected] (M.A.) 5 Institute of Organic Chemistry, Faculty of Chemistry, Lodz University of Technology, Zeromskiego˙ 116, 90-924 Lodz, Poland; [email protected] (K.K.); [email protected] (J.Z.) 6 NewDrug, Patras Science Park, 26500 Patras, Greece; [email protected] 7 Department of Physiology and Pharmacology,
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2011/0082079 A1 Spetzler Et Al
    US 2011 0082079A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0082079 A1 Spetzler et al. (43) Pub. Date: Apr. 7, 2011 (54) GLUCAGON-LIKE PEPTDE-1 DERVATIVES Publication Classification AND THEIR PHARMACEUTICAL USE (51) Int. Cl. A638/22 (2006.01) (75) Inventors: Jane Spetzler, Bronsho (DK); C07K I4/575 (2006.01) Lauge Schäffer, Lyngby (DK); A6IP3/10 (2006.01) Jesper Lau, Farum (DK); Thomas A6IP 9/12 (2006.01) Kruse, Herlev (DK); Patrick A6IP3/04 (2006.01) William Garibay, Holte (DK); A6IP 9/10 (2006.01) Steffen Runge, Frederiksberg A6IPI/00 (2006.01) (DK); Henning Thogersen, Farum A6IPL/04 (2006.01) (DK); Ingrid Petersson, Frederiksberg (DK) (52) U.S. Cl. .......................... 514/7.2:530/399; 514/11.7 (57) ABSTRACT (73) Assignee: Novo Nordisk A/S, Bagsvard (DK) The invention relates to protracted Glucagon-Like Peptide-1 Appl. No.: (GLP-1) derivatives and therapeutic uses thereof. The GLP-1 (21) 12/676,451 derivative of the invention comprises a modified GLP-1 (7- PCT Fled: 37) sequence having a total of 2-12 amino acid modifications, (22) Sep. 5, 2008 including Glu22 and Arg26, and being derivatised with an PCT NO.: PCT/EP2008/061755 albumin binding residue or pegylated in position 18, 20, 23. (86) 30, 31, 34, 36, 37, or 39. These compounds are useful in the S371 (c)(1), treatment or prevention of diabetes type 2 and related dis (2), (4) Date: Oct. 13, 2010 eases. The compounds are potent, stable, have long half-lives, a high affinity of binding to albumin, and/or a high affinity of (30) Foreign Application Priority Data binding to the extracellular domain of the GLP-1 receptor (GLP-1R), all of which is of potential relevance for the overall Sep.
    [Show full text]
  • PYY(3-36) Analogues: Structure
    PYY(3-36) analogues: Structure- activity relationships in energy homeostasis A thesis submitted for the degree of Doctor of Philosophy, Imperial College London 2011 Iain David Pritchard Section of Investigative Medicine Division of Diabetes, Endocrinology and Metabolism Department of Medicine Imperial College London Abstract The developed world is currently in the grip of an obesity epidemic. As a result, there is much ongoing research into the development of an effective anti-obesity agent. Peptide YY (PYY) is a 36 amino acid gastro-intestinal hormone released post- prandially by L-cells in the gastro-intestinal tract in proportion to the calorie content of a meal. The predominant form of the hormone found in circulation is the truncated PYY(3-36). Administration of PYY(3-36) at physiological doses to humans has been shown to reduce food intake. However, due to enzymatic degradation these effects are short lived, reducing the hormone’s utility as an anti-obesity pharmaceutical agent. A series of analogues of PYY(3-36) were designed either with amino acid substitutions in specific parts of the peptide sequence and/or with chemical modifications to the native sequence with the aim of increasing resistance to enzymatic activity whilst retaining or even enhancing the peptide’s bioactivity. The analogues were tested for resistance to degradation by different proteolytic enzymes in comparison to natural PYY(3-36). Their affinity to the Y2 receptor, for which PYY(3-36) is a natural agonist was then investigated. Finally, the effects of peripheral administration of selected analogues on food intake in overnight fasted mice were investigated.
    [Show full text]
  • Peptides As Therapeutics with Enhanced Bioactivity
    Send Orders of Reprints at [email protected] Current Medicinal Chemistry, 2012, 19, 4451-4461 4451 Peptides As Therapeutics with Enhanced Bioactivity D. Goodwin1, P. Simerska1 and I. Toth*,1,2, 1The University of Queensland, School of Chemistry and Molecular Biosciences; 2School of Pharmacy, St. Lucia 4072, Queensland, Australia Abstract: The development of techniques for efficient peptide production renewed interest in peptides as therapeutics. Numerous modi- fications for improving stability, transport and affinity profiles now exist. Several new adjuvant and carrier systems have also been developed, enhancing the immunogenicity of peptides thus allowing their development as vaccines. This review describes the established and experimental approaches for manufacturing peptide drugs and highlights the techniques currently used for improving their drug like properties. Keywords: Peptide, drug delivery, vaccine, manufacture, bioavailability, peptide therapeutic, immunogenicity, peptide drug, peptide synthe- sis, clinical trials. INTRODUCTION side-chain reactivity, degree of modification, incorporation of un- natural components, in addition to the required purity, solubility, Natural and synthetic peptides have shown promise as pharma- stability and scale. There are two strategies for peptide production - ceutics with the potential to treat a wide variety of diseases. This chemical synthesis and biological manufacturing. potential is often overshadowed by the inability of the peptides to reach their targets in an active form in vivo. The delivery of active Chemical Peptide Synthesis peptides is challenging due to inadequate absorption through the Chemical synthesis has been used for the production of peptides mucosa and rapid breakdown by proteolytic enzymes. Peptides are in both research and industry and led to the development of the usually selective and efficacious, therefore need only be present in majority of peptide drugs [2].
    [Show full text]
  • 1 Peptide Therapeutics
    Universal Free E-Book Store Donate Us, In order to keep our Service Alive, We have to pay for placing files (Abstracts, Books, Literature & Software) on File Hosting Servers, Your donations will make our process of payment a bit easier, Please use any one of the Payment Gateway for Donation. Never matter what amount you donate (10’s or 100’s or 1000’s). Universal Free E-Book Store Universal Free E-Book Store PEPTIDE CHEMISTRY AND DRUG DESIGN Universal Free E-Book Store Universal Free E-Book Store PEPTIDE CHEMISTRY AND DRUG DESIGN Edited by BEN M. DUNN Universal Free E-Book Store Copyright © 2015 by John Wiley & Sons, Inc. All rights reserved Published by John Wiley & Sons, Inc., Hoboken, New Jersey Published simultaneously in Canada No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, except as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization through payment of the appropriate per-copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750-8400, fax (978) 750-4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, (201) 748-6011, fax (201) 748-6008, or online at http://www.wiley.com/go/permissions. Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose.
    [Show full text]
  • Conformations of Heterochiral and Homochiral Proline-Pseudoproline
    Conformations of Heterochiral and Homochiral Proline-Pseudoproline ConformationsSegments in of Peptides: Heterochiral Context and Dependent HomochiralCis–Trans Proline-PseudoprolinePeptide SegmentsBond Isomerization in Peptides: Context Dependent Cis–Trans Peptide Bond Isomerization Kantharaju,1 Srinivasarao Raghothama,2 Upadhyayula Surya Raghavender,3 Subrayashastry Aravinda,3 Narayanaswamy Shamala,3 Padmanabhan Balaram1 1 Molecular Biophysics Unit, Indian Institute of Science, Bangalore, Karnataka 560012, India 2 NMR Research Centre, Indian Institute of Science, Bangalore, Karnataka 560012, India 3 Department of Physics, Indian Institute of Science, Bangalore, Karnataka 560012, India Received 9 March 2009; accepted 7 April 2009 Published online 16 April 2009 in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/bip.21207 ABSTRACT: stabilized by two intramolecular hydrogen bonds. Further truncation of the sequence gives an appreciable rise in the The pseudoproline residue (CPro, L-2,2-dimethyl-1,3- D thiazolidine-4-carboxylic acid) has been introduced into population of cis conformers in the tripeptide Piv- Pro- C heterochiral diproline segments that have been previously Pro-Leu-OMe (6). In the homochiral segment Piv-Pro- C shown to facilitate the formation of b-hairpins, containing Pro-Leu-OMe (7)onlythecis form is observed with the b central two and three residue turns. NMR studies of the NMR evidence strongly supporting a type VIa -turn ? octapeptide Boc-Leu-Phe-Val-DPro-CPro-Leu-Phe-Val- conformation, stabilized by a 4 1 hydrogen bond OMe (1), Boc-Leu-Val-Val-DPro-CPro-Leu-Val-Val-OMe between the Piv (CO) and Leu (3) NH groups. The crystal CH,CH3 (2), and the nonapeptide sequence Boc-Leu-Phe- structure of the analog peptide 7a (Piv-Pro- Pro- Val-DPro-CPro-DAla-Leu-Phe-Val-OMe (3)established Leu-NHMe) confirms the cis peptide bond geometry for CH,CH3 well-registered b-hairpin structures in chloroform the Pro- Pro peptide bond, resulting in a type VIa b # solution, with the almost exclusive population of the trans -turn conformation.
    [Show full text]
  • Université Du Québec À Montréal
    UNIVERSITÉ DU QUÉBEC À MONTRÉAL CONVERSION CONFORMATIONNELLE SECONDAIRE ET QUATERNAIRE D'HORMONES PEPTIDIQUES: ÉQUILIBRE ENTRE FONCTION BIOLOGIQUE ET AUTO-ASSEMBLAGE PATHOLOGIQUE THÈSE PRÉSENTÉE COMME EXIGENCE PARTIELLE DU DOCTORAT EN BIOCHIMIE PAR NOÉ QUITTOT SEPTEMBRE 2020 UNIVERSITÉ DU QUÉBEC À MONTRÉAL Service des bibliothèques Avertissement La diffusion de cette thèse se fait dans le respect des droits de son auteur, qui a signé le formulaire Autorisation de reproduire et de diffuser un travail de recherche de cycles supérieurs (SDU-522 – Rév.10-2015). Cette autorisation stipule que «conformément à l’article 11 du Règlement no 8 des études de cycles supérieurs, [l’auteur] concède à l’Université du Québec à Montréal une licence non exclusive d’utilisation et de publication de la totalité ou d’une partie importante de [son] travail de recherche pour des fins pédagogiques et non commerciales. Plus précisément, [l’auteur] autorise l’Université du Québec à Montréal à reproduire, diffuser, prêter, distribuer ou vendre des copies de [son] travail de recherche à des fins non commerciales sur quelque support que ce soit, y compris l’Internet. Cette licence et cette autorisation n’entraînent pas une renonciation de [la] part [de l’auteur] à [ses] droits moraux ni à [ses] droits de propriété intellectuelle. Sauf entente contraire, [l’auteur] conserve la liberté de diffuser et de commercialiser ou non ce travail dont [il] possède un exemplaire.» REMERCIEMENTS Au Professeur Steve Bourgault, à qui je souhaite tout d’abord exprimer ma gratitude pour m’avoir accepté en tant qu’étudiant au doctorat au sein de son laboratoire. Merci d’avoir été un directeur de thèse présent et à l’écoute tout au long de mon doctorat.
    [Show full text]
  • Peptide Publications from Mimotopes Pty Ltd Or Collaborators
    Peptide Publications 1 PEPTIDE PUBLICATIONS 1. Peptide publications from Mimotopes Pty Ltd or Collaborators Thomas, D. A.; Francis, P.; Smith, C.; Ratcliffe, S.; Ede, N. J.; Kay, C.; Wayne, G.; Martin, S. L.; Moore, K.; Amour, A.; Hooper, N. M., A broad-spectrum fluorescence-based peptide library for the rapid identification of protease substrates. Proteomics 2006, 6, (7), 2112-20. Ercole, F.; FitzGerald, M.; Perera, S.; Ede, N.; Riley, P.; Campbell, R., A combinatorial approach in designing hydrophilic surfaces for solid-phase peptide synthesis. J. Appl. Polym. Sci. 2003, 89, (12), 3371-3378. Khammanivong, V.; Liu, X. S.; Liu, W. J.; Rodda, S. J.; Leggatt, G. R.; Tindle, R. W.; Frazer, I. H.; Fernando, G. J., Paucity of functional CTL epitopes in the E7 oncoprotein of cervical cancer associated human papillomavirus type 16. Immunol. Cell Biol. 2003, 81, (1), 1-7. Nelson, P. N.; Westwood, O. M.; Soltys, A.; Jefferis, R.; Goodall, M.; Baumforth, K. R.; Frampton, G.; Tribbick, G.; Roden, D.; Hay, F. C., Characterisation of epitopes of pan- IgG/anti-G3m(u) and anti-Fc monoclonal antibodies. Immunol. Lett. 2003, 88, (1), 77-83. Takahashi, T.; Nagamiya, H.; Doi, T.; Griffiths, P. G.; Bray, A. M., Solid phase library synthesis of cyclic depsipeptides: aurilide and aurilide analogues. J. Comb. Chem. 2003, 5, (4), 414-428. Deliyannis, G.; Jackson, D. C.; Ede, N. J.; Zeng, W.; Hourdakis, I.; Sakabetis, E.; Brown, L. E., Induction of long-term memory CD8(+) T cells for recall of viral clearing responses against influenza virus. J. Virol. 2002, 76, (9), 4212-21. Ede, N.
    [Show full text]
  • Bachem ÂŒ Insights Into Peptide Chemistry Achievements by The
    874 CHIMIA 2013, 67, Nr. 12 PePtide Science in Switzerland doi:10.2533/chimia.2013.874 Chimia 67 (2013) 874–880 © Schweizerische Chemische Gesellschaft Bachem – Insights into Peptide Chemistry Achievements by the World’s Leading Independent Manufacturer of Peptides Monika Mergler, Günther Loidl, Martina Diekmann, and Fritz Dick* Abstract: The Swiss fine chemicals company Bachem, pioneer and specialist in the chemical synthesis of peptides, has also become an internationally leading manufacturer of peptide active pharmaceutical ingredients (APIs). In response to increasing demands in scale and purity, Bachem’s research efforts centered on the chemistry involved in solid-phase peptide synthesis and the production of the required amino acid derivatives with the aim of a continuous improvement of the technology. The resulting optimized protocols together with high-throughput equipment enabled us to manufacture long peptide APIs and, more recently, even pharma-grade glycoproteins in industrial scale. Keywords: Active pharmaceutical ingredients · Large-scale peptide manufacturing · Process development · Side reactions · Solid-phase peptide synthesis Introduction Bachem and is still in our portfolio of more Peptide Synthesis at Bachem than 50 generic APIs. More than 40 years of experience al- This achievement triggered a further The ‘classical’ solution synthesis still lows us to highlight research and develop- significant growth: Processes for manu- has its place at Bachem for manufactur- ment activities in the field of peptide chem- facturing of luteinizing hormone-releasing ing short peptides for research purposes istry by Bachem. Achievements in the field hormone (LHRH), LHRH agonists, and or, adhering to cGMP guidelines, for ge- of liquid- and solid-phase synthesis, with even large peptides such as salmon calci- neric APIs.
    [Show full text]
  • Chemical Methods for Peptide and Protein Production
    Molecules 2013, 18, 4373-4388; doi:10.3390/molecules18044373 OPEN ACCESS molecules ISSN 1420-3049 www.mdpi.com/journal/molecules Review Chemical Methods for Peptide and Protein Production Saranya Chandrudu 1, Pavla Simerska 1,* and Istvan Toth 1,2 1 School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, Qld 4072, Australia; E-Mails: [email protected] (S.C.); [email protected] (I.T.) 2 School of Pharmacy, The University of Queensland, St. Lucia, Qld 4072, Australia * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +61-7-3365-4636; Fax: +61-7-3365-4273. Received: 12 March 2013; in revised form: 28 March 2013 / Accepted: 9 April 2013 / Published: 12 April 2013 Abstract: Since the invention of solid phase synthetic methods by Merrifield in 1963, the number of research groups focusing on peptide synthesis has grown exponentially. However, the original step-by-step synthesis had limitations: the purity of the final product decreased with the number of coupling steps. After the development of Boc and Fmoc protecting groups, novel amino acid protecting groups and new techniques were introduced to provide high quality and quantity peptide products. Fragment condensation was a popular method for peptide production in the 1980s, but unfortunately the rate of racemization and reaction difficulties proved less than ideal. Kent and co-workers revolutionized peptide coupling by introducing the chemoselective reaction of unprotected peptides, called native chemical ligation. Subsequently, research has focused on the development of novel ligating techniques including the famous click reaction, ligation of peptide hydrazides, and the recently reported -ketoacid-hydroxylamine ligations with 5- oxaproline.
    [Show full text]
  • A Snake Toxin As a Theranostic Agent for the Type 2 Vasopressin Receptor
    A snake toxin as a theranostic agent for the type 2 vasopressin receptor Laura Droctové, Manon Lancien, Vu Long Tran, Michaël Susset, Benoit Jego, Frederic Theodoro, Pascal Kessler, Gilles Mourier, Philippe Robin, Sékou Siramakan Diarra, et al. To cite this version: Laura Droctové, Manon Lancien, Vu Long Tran, Michaël Susset, Benoit Jego, et al.. A snake toxin as a theranostic agent for the type 2 vasopressin receptor. Theranostics, Ivyspring International Publisher, 2020. hal-03070378 HAL Id: hal-03070378 https://hal.archives-ouvertes.fr/hal-03070378 Submitted on 15 Dec 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Theranostics 2020, Vol. 10, Issue 25 11580 Ivyspring International Publisher Theranostics 2020; 10(25): 11580-11594. doi: 10.7150/thno.47485 Research Paper A snake toxin as a theranostic agent for the type 2 vasopressin receptor Laura Droctové1, Manon Lancien1, Vu Long Tran2, Michaël Susset1, Benoit Jego2, Frederic Theodoro3, Pascal Kessler1, Gilles Mourier1, Philippe Robin1, Sékou Siramakan Diarra4, Stefano Palea4, Adrien Flahault5, Amélia Chorfa6, Maithé Corbani6, Catherine Llorens-Cortes7, Bernard Mouillac6, Christiane Mendre6, Alain Pruvost3, Denis Servent1, Charles Truillet2, Nicolas Gilles1 1. Université Paris Saclay, CEA, INRAE, Département Médicaments et Technologies pour la Santé (DMTS), SIMoS, 91191 Gif-sur-Yvette, France.
    [Show full text]
  • Synthesis of Sulfotyrosine Bearing Peptides and Analogues
    Synthesis of Sulfotyrosine Bearing Peptides and Analogues by Ahmed Magdy Ali A thesis presented to the University of Waterloo in fulfillment of the thesis requirement for the degree of Doctor of Philosophy in Chemistry Waterloo, Ontario, Canada, 2010 © Ahmed Magdy Ali 2010 AUTHOR'S DECLARATION I hereby declare that I am the sole author of this thesis. This is a true copy of the thesis, including any required final revisions, as accepted by my examiners. I understand that my thesis may be made electronically available to the public. ii Abstract Sulfation of tyrosine residues is a post-translational modification that occurs on many secretory as well as transmembrane proteins. This modification is believed to be essential for numerous biological processes. However, one of the factors hindering the study of the significance of sulfotyrosine (sTyr) within a protein is the absence of a general method that enables the synthesis of sTyr peptides in satisfactory yields and purity. A general approach to the synthesis of sTyr-bearing peptides was developed in which the sTyr residue is incorporated into the peptides with the sulfate group protected. For the implementation of this general approach a new protecting group for sulfates, namely, the dichlorovinyl (DCV) group was developed. This was accomplished by conducting a careful analysis of the reaction of a trichloroethyl (TCE)-protected sulfate ester with piperidine and 2-methylpiperidine (2-MP). A unique sulfuryl imidazolium reagent, compound 2.22, was also developed that enabled the ready synthesis of DCV-protected sulfates. This reagent was used to prepare the amino acid building block FmocTyr(SO3DCV)OH (2.23).
    [Show full text]