Towards a Biochemical Reconstitution of Nepenthes Pitcher Fluid for the Treatment of Celiac Disease

Total Page:16

File Type:pdf, Size:1020Kb

Towards a Biochemical Reconstitution of Nepenthes Pitcher Fluid for the Treatment of Celiac Disease University of Calgary PRISM: University of Calgary's Digital Repository Graduate Studies The Vault: Electronic Theses and Dissertations 2014-09-12 Towards a Biochemical Reconstitution of Nepenthes Pitcher Fluid for the Treatment of Celiac Disease Yang, Menglin Yang, M. (2014). Towards a Biochemical Reconstitution of Nepenthes Pitcher Fluid for the Treatment of Celiac Disease (Unpublished master's thesis). University of Calgary, Calgary, AB. doi:10.11575/PRISM/28475 http://hdl.handle.net/11023/1740 master thesis University of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission. Downloaded from PRISM: https://prism.ucalgary.ca UNIVERSITY OF CALGARY Towards a Biochemical Reconstitution of Nepenthes Pitcher Fluid for the Treatment of Celiac Disease by Menglin Yang A THESIS SUBMITTED TO THE FACULTY OF GRADUATE STUDIES IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE GRADUATE PROGRAM IN BIOCHEMISTRY AND MOLECULAR BIOLOGY CALGARY, ALBERTA SEPTEMBER, 2014 © Menglin Yang 2014 Abstract Celiac disease (CD) is an autoimmune disorder that is triggered by the incomplete digestion of gliadins in dietary gluten due to the abundance of P and Q residues in their protein sequence(s). This thesis provides an initial assessment of the proteolytic activity of Nepenthes plant extracts, which is attributed to the aspartic proteases, nepenthesin I and II, potential as an oral protease therapeutic for CD. To this end, nepenthesin I and II were produced recombinantly and characterized. The recombinant nepenthesins were able to reconstitute the proteolytic activity of Nepenthes extracts except for cleavage after P, which was attributed to a previously unidentified protease. Nevertheless, the Nepenthes extracts and recombinant nepenthesin I/II were assessed for their capacity to detoxify gliadins. Although the recombinant nepenthesins alone did not appear sufficient, the Nepenthes plant extracts appeared to efficiently detoxify gliadin, which supports the proposed formulations potential as an effective oral therapeutic for CD. ii Preface Portions of this thesis resulted in the following peer-reviewed or in preparation publications: 1) Rey, M., Yang, M., Burns, K.M., Yu, Y., Lees-Miller, S.P., and Schriemer, D.C. (2013). Nepenthesin from monkey cups for hydrogen/deuterium exchange mass spectrometry. Molecular and Cellular Proteomics, 12(2), 464-472 2) Yang, M., Hoeppner, M., Rey, M., Man, P. and Schriemer, D.C. (2014). Recombinant nepenthesin II for hydrogen/deuterium exchange mass spectrometry. In preparation. iii Acknowledgements First and foremost, I would like to thank my supervisor, Dr. David Schriemer, for giving me the opportunity to work on this project and for his immense support throughout my graduate studies. For their various experimental contributions to this project, I would like to thank Drs. Martial Rey, Laurent Brechenmacher and Kelvin Ma, as well as, Ronghua Yu. Thank you to my committee members, Drs. Hans Vogel and Tony Schryvers, for their advice and guidance throughout my graduate studies. Thank you to all of the members of the Schriemer laboratory for providing a great work environment. Thank you to my mother and father, Lanlan and Shanning, for their support in my career decisions. Finally, thank you to Patricia Lan for your unwavering support in all of my decisions. iv Dedication To Patti. v Table of Contents Abstract ........................................................................................................................................... ii Preface ............................................................................................................................................ iii Acknowledgements ........................................................................................................................ iv Dedication ....................................................................................................................................... v Table of Contents ........................................................................................................................... vi List of Tables ................................................................................................................................. ix List of Figures and Illustrations ...................................................................................................... x List of Symbols, Abbreviations, and Nomenclature .................................................................... xiii CHAPTER ONE: INTRODUCTION ............................................................................................. 1 1.1 Celiac Disease: A General Overview .................................................................................. 1 1.2 Epidemiology ....................................................................................................................... 1 1.3 Basis for Disease Development ........................................................................................... 3 1.4 Pathogenic Mechanism ........................................................................................................ 3 1.5 Disease Symptoms and Outcomes ....................................................................................... 7 1.6 Treatments for CD- The Gluten-Free Diet ........................................................................... 8 1.7 Proposed Therapeutics for CD ............................................................................................. 9 1.8 Oral Proteases: A Promising Treatment for CD ................................................................ 10 1.8.1 Oral Proteases: AlV003 ............................................................................................. 11 1.8.2 Oral Proteases: STAN1 .............................................................................................. 12 1.8.3 Oral Proteases: AN-PEP ............................................................................................ 13 1.8.4 Oral Proteases: Limitations and Future Directions .................................................... 13 1.9 A New Oral Protease Candidate for the Treatment of CD- Nepenthesin .......................... 14 1.10 Research Hypothesis and Objectives ............................................................................... 18 CHAPTER TWO: IDENTIFICATION OF THE PROTEOLYTIC COMPONENTS OF NEPENTHES PITCHER FLUID .................................................................................................. 19 2.1 Introduction ........................................................................................................................ 19 2.2 Experimental Procedures ................................................................................................... 20 2.2.1 Chemicals ................................................................................................................... 20 2.2.2 Horticulture of Nepenthes Plants ............................................................................... 20 2.2.3 Preparation of Nepenthes fluid for Proteome Studies ................................................ 22 2.2.4 Proteome Mass Spectrometry and Data Analysis ...................................................... 22 2.2.5 Visualization of the Nepenthes fluid Proteome over Time ........................................ 23 2.2.6 In-gel Processing ........................................................................................................ 23 2.2.7 Activity Assays .......................................................................................................... 24 2.2.8 Pepstatin A Purification ............................................................................................. 24 2.2.9 Determination of Cleavage Specificities ........................................................................ 25 2.3 Results and Discussion ...................................................................................................... 26 2.3.1 In-solution Proteome Analyses of Nepenthes Pitcher Secretions .............................. 26 2.3.2 Activity and Cleavage Preferences of Nepenthes Pitcher Fluid ................................ 34 2.3.3 Pepstatin A Purified Nepenthes Pitcher Fluid ............................................................ 39 2.4 Conclusions ........................................................................................................................ 41 2.5 Contributions to the Chapter .............................................................................................. 41 vi CHAPTER THREE: RECOMBINANT RECONSTITUTION OF THE PROTEOLYTIC ACTIVITY OF NEPENTHES PITCHER FLUID ........................................................................ 42 3.1 Introduction ........................................................................................................................ 42 3.2 Experimental Procedures ................................................................................................... 43 3.2.1. Chemicals .................................................................................................................. 43 3.2.2. Plasmid Preparation .................................................................................................
Recommended publications
  • Genome Skimming Provides Well Resolved Plastid and Nuclear
    Australian Systematic Botany, 2019, 32, 243–254 ©CSIRO 2019 https://doi.org/10.1071/SB18057 Supplementary material Genome skimming provides well resolved plastid and nuclear phylogenies, showing patterns of deep reticulate evolution in the tropical carnivorous plant genus Nepenthes (Caryophyllales) Lars NauheimerA,B,C,G, Lujing CuiD,E, Charles ClarkeA, Darren M. CraynA,B,C,D, Greg BourkeF and Katharina NargarA,B,C,D AAustralian Tropical Herbarium, James Cook University, PO Box 6811, Cairns, Qld 4878, Australia. BCentre for Tropical Environmental Sustainability Science, James Cook University, McGregor Road, Smithfield, Qld 4878, Australia. CCentre for Tropical Bioinformatics and Molecular Biology, James Cook University, McGregor Road, Smithfield, Qld 4878, Australia. DNational Research Collections Australia, Commonwealth Industrial and Scientific Research Organisation (CSIRO), GPO Box 1700, Canberra, ACT 2601, Australia. ESchool of Computer Science and Engineering, University of New South Wales, NSW 2052, Australia. FBlue Mountains Botanic Garden, Bells Line of Road, Mount Tomah, NSW 2758, Australia. GCorresponding author. Email: lars.nauheimer@jcu.edu.au Page 1 of 6 Australian Systematic Botany ©CSIRO 2019 https://doi.org/10.1071/SB18057 Table S1. List of accessions used for phylogenetic analyses with sectional association, voucher number, geographic origin and DNA number All herbarium vouchers are located in the Australian Tropical Herbarium in Cairns (CNS) Species Section Voucher Origin DNA number Nepenthes ampullaria Jack Urceolatae Clarke, C. & Bourke, G. 2 Borneo, Malaysia G07903 Nepenthes benstonei C.Clarke Pyrophytae Clarke, C. & Bourke, G. 38 Malay Peninsula, Malaysia G07897 Nepenthes bokorensis Mey × Nepenthes ventricosa Blanco Pyrophytae × Insignes Clarke, C. & Bourke, G. 54 Horticulatural G07899 Nepenthes bongso Korth. Montanae Clarke, C.
    [Show full text]
  • Membrane Proteins • Cofactors – Plimstex • Membranes • Dna • Small Molecules/Gas • Large Complexes
    Structural mass spectrometry hydrogen/deuterium exchange Petr Man Structural Biology and Cell Signalling Institute of Microbiology, Czech Academy of Sciences Structural biology methods Low-resolution methods High-resolution methods Rigid SAXS IR Raman CD ITC MST Cryo-EM AUC SPR MS X-ray crystallography Chemical cross-linking H/D exchange Native ESI + ion mobility Oxidative labelling Small Large NMR Dynamic Structural biology approaches Simple MS, quantitative MS Cross-linking, top-down, native MS+dissociation native MS+ion mobility Cross-linking Structural MS What can we get using mass spectrometry IM – ion mobility CXL – chemical cross-linking AP – afinity purification OFP – oxidative footprinting HDX – hydrogen/deuterium exchange ISOTOPE EXCHANGE IN PROTEINS 1H 2H 3H occurence [%] 99.988 0.0115 trace 5 …Kaj Ulrik Linderstrøm-Lang „Cartesian diver“ Proteins are migrating in tubes with density gradient until they stop at the point where the densities are equal 1H 2H 3H % 99.9885 0.0115 trace density [g/cm3] 1.000 1.106 1.215 Methods of detection IR: β-: NMR: 1 n = 1.6749 × 10-27 kg MS: 1H 2H 3H výskyt% [%] 99.9885 0.0115 trace hustotadensity vody [g/cm [g/cm3] 3] 1.000 1.106 1.215 jadernýspinspin ½+ 1+ ½+ mass [u] 1.00783 2.01410 3.01605 Factors affecting H/D exchange hydrogen bonding solvent accessibility Factors affecting H/D exchange Side chains (acidity, steric shielding) Bai et al.: Proteins (1993) Glasoe, Long: J. Phys. Chem. (1960) Factors affecting H/D exchange – side chain effects Inductive effect – electron density is Downward shift due to withdrawn from peptide steric hindrance effect of bond (S, O).
    [Show full text]
  • The First Record of the Boreal Bog Species Drosera Rotundifolia (Droseraceae) from the Philippines, and a Key to the Philippine Sundews
    Blumea 61, 2016: 24–28 www.ingentaconnect.com/content/nhn/blumea RESEARCH ARTICLE http://dx.doi.org/10.3767/000651916X691330 The first record of the boreal bog species Drosera rotundifolia (Droseraceae) from the Philippines, and a key to the Philippine sundews F.P. Coritico1, A. Fleischmann2 Key words Abstract Drosera rotundifolia, a species of the temperate Northern Hemisphere with a disjunct occurrence in high montane West Papua, has been discovered in a highland peat bog on Mt Limbawon, Pantaron Range, Bukidnon carnivorous plants on the island of Mindanao, Philippines, which mediates to the only other known tropical, Southern Hemisphere Drosera location in New Guinea and the closest known northern populations in southern Japan and south-eastern China. Droseraceae A dichotomous key to the seven Drosera species of the Philippines is given, and distribution maps are provided. Malesia Mindanao Published on 15 March 2016 Northern Hemisphere - Tropics disjunction Philippines INTRODUCTION Drosera rotundifolia L. (the generic type) is a temperate, winter dormant species that is widespread in the Northern The Philippines are rich in carnivorous plants, with about 47 Hemisphere, from Pacific North America across large parts of species known from the islands, most of which belong to the northern America and Europe to Siberia and the Kamchatka pitcher plant genus Nepenthes L. This genus has more than 30 Peninsula, South Korea and Japan. It is the Drosera spe- species in the Philippines, all except Nepenthes mirabilis (Lour.) cies covering the largest range, spanning the entire Northern Druce endemic to the country. Most species occur on Mindanao Hemisphere from 180° Western Longitude to about 180° East, and Palawan, while several are confined to a single highland however, not forming a continuous circumboreal range (Diels or even mountain peak (Robinson et al.
    [Show full text]
  • Discovery of Digestive Enzymes in Carnivorous Plants with Focus on Proteases
    A peer-reviewed version of this preprint was published in PeerJ on 5 June 2018. View the peer-reviewed version (peerj.com/articles/4914), which is the preferred citable publication unless you specifically need to cite this preprint. Ravee R, Mohd Salleh F‘, Goh H. 2018. Discovery of digestive enzymes in carnivorous plants with focus on proteases. PeerJ 6:e4914 https://doi.org/10.7717/peerj.4914 Discovery of digestive enzymes in carnivorous plants with focus on proteases Rishiesvari Ravee 1 , Faris ‘Imadi Mohd Salleh 1 , Hoe-Han Goh Corresp. 1 1 Institute of Systems Biology (INBIOSIS), Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia Corresponding Author: Hoe-Han Goh Email address: gohhh@ukm.edu.my Background. Carnivorous plants have been fascinating researchers with their unique characters and bioinspired applications. These include medicinal trait of some carnivorous plants with potentials for pharmaceutical industry. Methods. This review will cover recent progress based on current studies on digestive enzymes secreted by different genera of carnivorous plants: Drosera (sundews), Dionaea (Venus flytrap), Nepenthes (tropical pitcher plants), Sarracenia (North American pitcher plants), Cephalotus (Australian pitcher plants), Genlisea (corkscrew plants), and Utricularia (bladderworts). Results. Since the discovery of secreted protease nepenthesin in Nepenthes pitcher, digestive enzymes from carnivorous plants have been the focus of many studies. Recent genomics approaches have accelerated digestive enzyme discovery. Furthermore, the advancement in recombinant technology and protein purification helped in the identification and characterisation of enzymes in carnivorous plants. Discussion. These different aspects will be described and discussed in this review with focus on the role of secreted plant proteases and their potential industrial applications.
    [Show full text]
  • Recircumscription of the Nepenthes Alata Group (Caryophyllales: Nepenthaceae), in the Philippines, with Four New Species
    European Journal of Taxonomy 69: 1-23 ISSN 2118-9773 http://dx.doi.org/10.5852/ejt.2013.69 www.europeanjournaloftaxonomy.eu 2013 · Martin Cheek & Matthew Jebb This work is licensed under a Creative Commons Attribution 3.0 License. Research article Recircumscription of the Nepenthes alata group (Caryophyllales: Nepenthaceae), in the Philippines, with four new species Martin CHEEK1 & Matthew JEBB2 1 Herbarium, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3AE, U.K. Email: m.cheek@kew.org (corresponding author) 2 National Botanic Garden, Glasnevin, Dublin 9, Ireland Email: matthew.jebb@opw.ie Abstract. An overview of Nepenthes in the Philippines is presented. Four new species, Nepenthes extincta sp. nov., N. kitanglad sp. nov., N. kurata sp. nov. and N. leyte sp. nov. are described and illustrated from the Philippines and placed in the Nepenthes alata group. An updated circumscription and key to the species of the group is provided. Delimitation and comparison with the Regiae group is given. All four of the newly described species are assessed as threatened using the International Union for the Conservation of Nature 2012 standard, and one, N. extincta sp. nov. is considered likely to be already extinct due to open-cast mining. Logging and conversion of forest habitat are thought to be the main threats to the other three species. Key words. Conservation, Nepenthes alata group, Mindanao, threatened, ultramafic. Cheek M. & Jebb M. 2013. Recircumscription of the Nepenthes alata group (Caryophyllales: Nepenthaceae), in the Philippines, with four new species. European Journal of Taxonomy 69: 1-23. http://dx.doi.org/10.5852/ ejt.2013.69 Introduction This paper forms part of studies towards a World Monograph of Nepenthes L.
    [Show full text]
  • Pricelist March 2019
    PRICELIST MARCH 2019 About us. Passionate about carnivorous plants from a young age, Scotland Carnivorous Plants was established in 2014 by myself, Oliver Murray. At Scotland Carnivorous Plants we specialise in the sale of the highest quality potted nepenthes. We strive for excellence and precision in every detail from plant health to customer service and packaging. We are one of the largest Borneo Exotics distributors in Europe, Importing since 2015. Please share my passion for nepenthes with me and do not hesitate to contact me, I am always willing to chat anything carnivorous plants! Please have a look at the reviews on our eBay page, we are sure you will not be disappointed. Ordering from us Here are some quick details about ordering from us… o All plants are sent potted unless otherwise stated. o Plants are wrapped in the highest quality materials protected for winter, with thermally insulated packaging - heatpacks available. o All plants are sent with appropriate plant passport documentation. o Guaranteed safe arrival and the highest quality. (Europe only!). o Please contact us, to place your order. o Photos of plants provided on request. o TRADES welcome: I am always happy to trade, contact me. o Photos on this pricelist are largely supplied from Borneo exotics and give an indication of what plants will grow to look like. o Pre-orders, we offer plants due to arrive in out next shipment (end of April/early May), these can be sent to you the day we receive them, or we can acclimate free of charge. o Payment is with PayPal (3.5% of total bill service charge).
    [Show full text]
  • Serine Proteases with Altered Sensitivity to Activity-Modulating
    (19) & (11) EP 2 045 321 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 08.04.2009 Bulletin 2009/15 C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12Q 1/37 (2006.01) (21) Application number: 09150549.5 (22) Date of filing: 26.05.2006 (84) Designated Contracting States: • Haupts, Ulrich AT BE BG CH CY CZ DE DK EE ES FI FR GB GR 51519 Odenthal (DE) HU IE IS IT LI LT LU LV MC NL PL PT RO SE SI • Coco, Wayne SK TR 50737 Köln (DE) •Tebbe, Jan (30) Priority: 27.05.2005 EP 05104543 50733 Köln (DE) • Votsmeier, Christian (62) Document number(s) of the earlier application(s) in 50259 Pulheim (DE) accordance with Art. 76 EPC: • Scheidig, Andreas 06763303.2 / 1 883 696 50823 Köln (DE) (71) Applicant: Direvo Biotech AG (74) Representative: von Kreisler Selting Werner 50829 Köln (DE) Patentanwälte P.O. Box 10 22 41 (72) Inventors: 50462 Köln (DE) • Koltermann, André 82057 Icking (DE) Remarks: • Kettling, Ulrich This application was filed on 14-01-2009 as a 81477 München (DE) divisional application to the application mentioned under INID code 62. (54) Serine proteases with altered sensitivity to activity-modulating substances (57) The present invention provides variants of ser- screening of the library in the presence of one or several ine proteases of the S1 class with altered sensitivity to activity-modulating substances, selection of variants with one or more activity-modulating substances. A method altered sensitivity to one or several activity-modulating for the generation of such proteases is disclosed, com- substances and isolation of those polynucleotide se- prising the provision of a protease library encoding poly- quences that encode for the selected variants.
    [Show full text]
  • Nepenthes Argentii Philippines, N. Aristo
    BLUMEA 42 (1997) 1-106 A skeletal revision of Nepenthes (Nepenthaceae) Matthew Jebb & Martin Chee k Summary A skeletal world revision of the genus is presented to accompany a family account forFlora Malesi- ana. 82 species are recognised, of which 74 occur in the Malesiana region. Six species are described is raised from and five restored from as new, one species infraspecific status, species are synonymy. Many names are typified for the first time. Three widespread, or locally abundant hybrids are also included. Full descriptions are given for new (6) or recircumscribed (7) species, and emended descrip- Critical for all the Little tions of species are given where necessary (9). notes are given species. known and excluded species are discussed. An index to all published species names and an index of exsiccatae is given. Introduction Macfarlane A world revision of Nepenthes was last undertaken by (1908), and a re- Malesiana the gional revision forthe Flora area (excluding Philippines) was completed of this is to a skeletal revision, cover- by Danser (1928). The purpose paper provide issues which would be in the ing relating to Nepenthes taxonomy inappropriate text of Flora Malesiana.For the majority of species, only the original citation and that in Danser (1928) and laterpublications is given, since Danser's (1928) work provides a thorough and accurate reference to all earlier literature. 74 species are recognised in the region, and three naturally occurring hybrids are also covered for the Flora account. The hybrids N. x hookeriana Lindl. and N. x tri- chocarpa Miq. are found in Sumatra, Peninsular Malaysia and Borneo, although rare within populations, their widespread distribution necessitates their inclusion in the and other and with the of Flora.
    [Show full text]
  • The Coordinate Regulation of Digestive Enzymes in the Pitchers of Nepenthes Ventricosa
    Rollins College Rollins Scholarship Online Honors Program Theses Spring 2020 The Coordinate Regulation of Digestive Enzymes in the Pitchers of Nepenthes ventricosa Zephyr Anne Lenninger zlenninger@rollins.edu Follow this and additional works at: https://scholarship.rollins.edu/honors Part of the Plant Biology Commons Recommended Citation Lenninger, Zephyr Anne, "The Coordinate Regulation of Digestive Enzymes in the Pitchers of Nepenthes ventricosa" (2020). Honors Program Theses. 120. https://scholarship.rollins.edu/honors/120 This Open Access is brought to you for free and open access by Rollins Scholarship Online. It has been accepted for inclusion in Honors Program Theses by an authorized administrator of Rollins Scholarship Online. For more information, please contact rwalton@rollins.edu. The Coordinate Regulation of Digestive Enzymes in the Pitchers of Nepenthes ventricosa Zephyr Lenninger Rollins College 2020 Abstract Many species of plants have adopted carnivory as a way to obtain supplementary nutrients from otherwise nutrient deficient environments. One such species, Nepenthes ventricosa, is characterized by a pitcher shaped passive trap. This trap is filled with a digestive fluid that consists of many different digestive enzymes, the majority of which seem to have been recruited from pathogen resistance systems. The present study attempted to determine whether the introduction of a prey stimulus will coordinately upregulate the enzymatic expression of a chitinase and a protease while also identifying specific chitinases that are expressed by Nepenthes ventricosa. We were able to successfully clone NrCHIT1 from a mature Nepenthes ventricosa pitcher via a TOPO-vector system. In order to asses enzymatic expression, we utilized RT-qPCR on pitchers treated with chitin, BSA, or water.
    [Show full text]
  • ( 12 ) United States Patent
    US009745565B2 (12 ) United States Patent (10 ) Patent No. : US 9 , 745 , 565 B2 Schriemer (45 ) Date of Patent: * Aug. 29 , 2017 ( 54 ) TREATMENT OF GLUTEN INTOLERANCE FOREIGN PATENT DOCUMENTS AND RELATED CONDITIONS EP 2 090 662 A2 8 /2009 WO WO - 2010 /021752 2 / 2010 ( 71 ) Applicant: Nepety , LLC , Destin , FL (US ) WO WO - 2011 /097266 8 / 2011 ( 72 ) Inventor: David Schriemer , Chestermere (CA ) WO WO - 2011 / 126873 10 / 2011 ( 73 ) Assignee : NEPETX , LLC , Destin , FL (US ) OTHER PUBLICATIONS Adlassnig W et al . Traps of carnivorous pitcher plants as a habitat: ( * ) Notice : Subject to any disclaimer, the term of this composition of the fluid , biodiversity and mutualistic activities . patent is extended or adjusted under 35 2011. Annals of Botany . 107 : 181 - 194 . * U . S . C . 154 (b ) by 98 days . Amagase , et al . , “ Acid Protease in Nepenthes, " The Journal of Biochemistry , ( 1969 ) , 66 ( 4 ) :431 -439 . This patent is subject to a terminal dis Athauda , et al . “ Enzymic and structural characterization of claimer . nepenthesin , a unique member of a novel subfamily of aspartic proteinases , ” Biochemical Journal ( 2004 ) 381 ( 1 ) :295 - 306 . (21 ) Appl. No. : 14/ 506 ,456 Bennett et al. , “ Discovery and Characterization of the Laulimalide Microtubule Binding Mode by Mass Shift Perturbation Mapping ," Chemistry & Biology , (2010 ) , 17 :725 - 734 . ( 22 ) Filed : Oct. 3 , 2014 Bethune, et al. , " Oral enzyme therapy for celiac sprue ,” Methods Enzymol. , (2012 ) , 502 :241 - 271 . (65 65) Prior Publication Data Blonder et al. , “ Proteomic investigation of natural killer cell US 2015 /0265686 A1 Sep . 24 , 2015 microsomes using gas -phase fractionation by mass spectrometry, " Biochimica et Biophysica Acta , (2004 ) , 1698 :87 -95 .
    [Show full text]
  • 20Th International Mass Spectrometry Conference
    IMSC 2014 20th International Mass Spectrometry Conference August 24-29, 2014 Geneva, Switzerland PROGRAM v. 17.09.2014 More targets. More accurately. Faster than ever. Analytical challenges grow in quantity and complexity. Quantify a larger number of compounds and more complex analytes faster and more accurately with our new portfolio of LC-MS instruments, sample prep solutions and software. High-resolution, accurate mass solutions using Thermo Scientific™ Orbitrap™ MS quantifies all detectable compounds with high specificity, and triple quadrupole MS delivers SRM sensitivity and speed to detect targeted compounds more quickly. Join us in meeting today’s challenges. Together we’ll transform quantitative science. Quantitation transformed. • Discover more at thermoscientific.com/quan-transformed • Visit thermoscientific.com/imsc or booth 23 for more information © 2014 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are © 2014 Thermo Fisher Scientific Inc. All rights the property of Thermo Fisher Scientific and its subsidiaries. the property Thermo Scientific™ Q Exactive™ HF MS Thermo Scientific™ TSQ Quantiva™ MS Thermo Scientific™ TSQ Endura™ MS Screen and quantify known and unknown targets Leading SRM sensitivity and speed Ultimate SRM quantitative value and with HRAM Orbitrap technology in a triple quadrupole MS/MS unprecedented usability TABLE OF CONTENTS th 1. Welcome from the Chairs of the 20 IMSC ............................................................................................................................
    [Show full text]
  • A Dictionary of the Plant Names of the Philippine Islands," by Elmer D
    4r^ ^\1 J- 1903.—No. 8. DEPARTMEl^T OF THE IE"TEIlIOIi BUREAU OF GOVERNMENT LABORATORIES. A DICTIONARY OF THE PLAIT NAMES PHILIPPINE ISLANDS. By ELMER D, MERRILL, BOTANIST. MANILA: BUREAU OP rUKLIC I'RIN'TING. 8966 1903. 1903.—No. 8. DEPARTMEE^T OF THE USTTERIOR. BUREAU OF GOVEENMENT LABOEATOEIES. r.RARV QaRDON A DICTIONARY OF THE PLANT PHILIPPINE ISLANDS. By ELMER D. MERRILL, BOTANIST. MANILA: BUREAU OF PUBLIC PRINTING. 1903. LETTEE OF TEANSMITTAL. Department of the Interior, Bureau of Government Laboratories, Office of the Superintendent of Laboratories, Manila, P. I. , September 22, 1903. Sir: I have the honor to submit herewith manuscript of a paper entitled "A dictionary of the plant names of the Philippine Islands," by Elmer D. Merrill, Botanist. I am, very respectfully. Paul C. Freer, Superintendent of Government Laboratories. Hon. James F. Smith, Acting Secretary of the Interior, Manila, P. I. 3 A DICTIONARY OF THE NATIVE PUNT NAMES OF THE PHILIPPINE ISLANDS. By Elmer D. ^Ikkrii.i., Botanist. INTRODUCTIOX. The preparation of the present work was undertaken at the request of Capt. G. P. Ahern, Chief of the Forestry Bureau, the objeet being to facihtate the work of the various employees of that Bureau in identifying the tree species of economic importance found in the Arcliipelago. For the interests of the Forestry Bureau the names of the va- rious tree species only are of importance, but in compiling this list all plant names avaliable have been included in order to make the present Avork more generally useful to those Americans resident in the Archipelago who are interested in the vegetation about them.
    [Show full text]