Sequences of amino acid residues in bacitracin PDF
Preview Sequences of amino acid residues in bacitracin
Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1954 Sequences of amino acid residues in bacitracin John F. Griffith Iowa State College Follow this and additional works at:https://lib.dr.iastate.edu/rtd Part of theBiochemistry Commons Recommended Citation Griffith, John F., "Sequences of amino acid residues in bacitracin " (1954).Retrospective Theses and Dissertations. 13086. https://lib.dr.iastate.edu/rtd/13086 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please [email protected]. NOTE TO USERS This reproduction is the best copy available. UMI BMmmmB m AMIMO AOID KSi ctus ii lACifMeii F. ti'lffitto A Mmert&tiou Siiteitt®d t& tli® Q-ra«€ a%'® Fmmlty In Pst»t iml ©f Tk® f©i* the Btfr#e of mQfm if PlILQSOPli Approfets Signature was redacted for privacy. In •efeai'g® ©r Mmt Signature was redacted for privacy. I@M of Major Depart®@,nt Signature was redacted for privacy. B#an -of /'G-raduat# Gell^«gs Xowa Sta%® Golleg® 1954 UMI Number: DP12304 INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. UMI UMI Microform DP12304 Copyright 2005 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 il OP 551 r~v 3% c. 1 Page IlfEODIIGTIOi 1 mttm Of fiE LifiSAfon 3 • Bacitr&eiii A $ Mmlm aelA ooiiipoiltl©n $ StaMlitr 7 fitrstiom feeimTior 8 fniiftloaal gro'tipi ' 9 Amino gPOttp® 9 Carbcdjl group® 11 The •tttll'Mr 11 Ultraviolet absorption spe®traa 14 loleeular weight 15 Affiiii© ®.eit rtiitee stqueEots 16 Otber Baeitr&0in Peptld## If Amino it©id eoinpoiitioii. If UltraTiol@t Absorption spectra 20. .E3g>lEIMEIfAL 22 Materials 22 ia«itracin 22 Chemicale safl reagenti 22 Bacterial ©ulturet 23 Metliodi 23 Mierobiologieal assay® 23 Otiroiiatofr&ptiio laentifioation© on pap@r 2® Aeeending teehnique ' 26 OtieeMlng tecimiqtie Zf Location of aalno aeidfl 27 Tll^73 ill c-uantitatlfe ©teroMst©gr&pfe|' 't>m®X"30 Study of carboxoid lileskiiig ©©aditione Sequence study I Sequence study K Rate study 1. Partial hydrolysis with hydrochlorie acit-aoetie aclA Partial hydrolysii Blocking treptment® Complete t^drolysig Rate Btudy II. fartl&l tiydrolysis witM hydrochloric aeid-acetie acid terminal residue study with recovery taaplee Blocking tr#at»snti Complete k^di^olysl® Preparation of desiilfwrized b&citraein Preparation ©f Raney nickel catalyst Hydrogsnolysie of bacitracin Qualitative analytis of product Stqmenee study of detulfurized bacitracin Blocking trt&tatntfi Complete hydrolysis Extraction ©f thlohydantoini ani. pheny1thiohyd&ntoins Alkaline l^ydrolysiB of extrseti Perfowi© acid oxidation ©f bacitracin A Beqmum study of oxidised bacitracin A l£S:0Lfi Nlnhydrin Color Yield® Carboxoid Blocking le&ction Condition# Bacitracin Sequenee Studies Bate Studies feroinal Residue .Stuiiee Bacitracin Desulfurieed bacitracin Oxidized bacitracin A Ir SISCUSSIOH Carboxoid Bl®©kifig Itaetlou- Gotttitlsnp Sequence Stwfilts laeltratla Rate Studies Terminal ReBidue Stttti®& Beeitracin Desulfurized 'bacitraein OaiitiasA feaeltr-scin 1 SffiEftBX ASB eiMGLCSlOSS LIfllAfWg i'lfi© MWOMmmmnm 1 mmommm leesnt years have produced many variation® and a ftw Iffiproveasenti In the methods for anino acid reildu© tequtne® analysis ©f peptldts. the us© ©f 2,^-dinltrofluorobenE©Et hy Sangtr and alk&lln® phtnyl liothlocyanat# by Edii&n art two txa»t)l©s of a«lii©id blocking reagenti whloh h&ve found wld« application. Most of the »®thodg of Bt<|uenee analyels are of a qualitative nature, used in eonjunotion with iuch tech- ni«|u@e a.B paper and colu»n chroaat©gra|>hy, paper el@etro- phortsle, and counter current dletrlhutlon. B©eauee th©e® art qualitative aethods, thi peptide under eonslderatlon must h© hoaogeneous, or at least free of other peptide material, so that other substancte present cannot eaus« misleading rtsulti. Unfortunately few, if any, of the naturally occur ring peptide# are hoaogeneous» so that ofttn laborious puri fication proceduree mufit be carried out Cs.g., oxytocin). In spite of thle, many natural peptides h&ve been purified, and soae, fluch am ©•raalcldln S, insulin, tyrocldln, oxytocin, and vasopretiln, have had their amino acid rtsldu® tequences elucidated. It has become clear that ftw of the naturally occurring peptide® have a einple, straight chain gtructure. iever&l have been shown to be of a cyclic nature, ©tpeciftlly those 2 haTing aatifeletlo Bropertits j. somt eontaln non-peptia® aoletle#, fhe preeenee of such cyolic arrangenenta of resi dues &M ©ther unorthodox linkages, togsther with tht diffi culty iii separating and purifying sliiilar peptide® has suggested that a quantitatif© approach to structure analysis might perait structure atteralnation of « com|>l« peptid# which ie the »«.Jor coaponent of a oixture. fh© antihiotic, Mcltraoin, is such a mixture of pep tide®. It ii a faoily of at Itaet six «i»ilar peptides with & oajor 0onpon®nt, haeitraein 1, providing woat of the anti- •felotic sctlTity. fht InTeetigatlon upon which this thesle ie based I0 eonetrned with dtteraining tht tequence of amino acid reslduts in feaeltraoin toy application of QuantltatlT© aethodi to the crude aaterial.
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