http://api.elsevier.com.ucc.idm.oclc.org/content/abstract/scopus_id/0037070197 SCOPUS_ID:0037070197 11852050 S0014579301033130 10.1016/S0014-5793(01)03313-0 The orientation of the antibiotic peptide maculatin 1.1 in DMPG and DMPC lipid bilayers. Support for a pore-forming mechanism Journal j 40 FEBS Letters 00145793 512 1-3 47 51 47-51 2002-02-13 C.S.B. Chia C.S.B. Chia C.S.B. Chia C. Chia C. S B http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/55126316600 Maculatin 1.1 is an antimicrobial peptide isolated from the Australian tree frog Litoria genimaculata that adopts an amphipathic, ?-helical structure in solution. Its orientation and conformation when incorporated to pre-formed DMPG (1,2-dimyristoyl-sn-glycero-3-phosphoglycerol) and DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) vesicles was determined using polarised Fourier transform infrared-attenuated total reflection infrared and deuterium exchange experiments. For DMPG membranes, our results show insertion of ?70% of the maculatin 1.1 molecules, with an angle of insertion of approximately 35° to the membrane normal and with a predominant ?-helical structure. These results suggest that maculatin 1.1 acts through a pore-forming mechanism to lyse bacterial membranes. A similar degree of insertion in DMPG (65%) and ?-helical structure was observed for a biologically inactive, less amphipathic maculatin 1.1 analogue, P15A, although the helix tilt was found to be greater (46°) than for maculatin 1.1. Similar experiments performed using DMPC liposomes showed poor insertion, less than 5%, for both maculatin 1.1 and its analogue. In addition, the shape of the amide I band in these samples is consistent with ?-helix, ?-structure and disordered structures being present in similar proportion. These results clearly show that maculatin 1.1 inserts preferentially in negatively charged membranes (DMPG) which mimic the negatively charged membrane of Gram-positive bacteria. We attribute the high percentage of insertion of the biologically inactive analogue in DMPG to the fact that its concentration on the membrane surface in our experiments is likely to be much higher than that found in physiological conditions. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. 1507371757 University of Adelaide Nanyang Technological University University of Cambridge C.S.B. Chia C.S.B. Chia C.S.B. Chia C. Chia C. S B http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/55126316600 J. Torres J. Torres J. Torres J. Torres J. http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/7402798449 M.A. Cooper M.A. Cooper M.A. Cooper M. Cooper M. A. http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/7404411451 I.T. Arkin I.T. Arkin I.T. Arkin I. Arkin I. T. http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/7003858738 J.H. Bowie J.H. Bowie J.H. Bowie J. Bowie J. H. http://api.elsevier.com.ucc.idm.oclc.org/content/author/author_id/7102792890 Biochemistry Biophysics Molecular Biology Copyright 2008 Elsevier B.V., All rights reserved. MEDLINE® is the source for the MeSH terms of this document. S0014579301033130 10.1016/S0014-5793(01)03313-0 34164449 2002074089 2002047749 11852050 0037070197 0037070197 EMBASE CABS MEDL Scopusbase 1,2-Dimyristoyl-sn-glycero-3-phosphocholine 1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol Attenuated total reflection Fourier transform infrared Litoria genimaculata Maculatin 1.1 Peptide antibiotic Phospholipid membrane The orientation of the antibiotic peptide maculatin 1.1 in DMPG and DMPC lipid bilayers. Support for a pore-forming mechanism C.S.B. Chia C.S.B. Chia C.S.B. Chia C. Chia C. S B M.A. Cooper M.A. Cooper M.A. Cooper M. Cooper M. A. Department of Chemistry University of Cambridge Lensfield Road Cambridge CB2 1EW J. Torres J. Torres J. Torres J. Torres J. jtorres@ntu.edu.sg I.T. Arkin I.T. Arkin I.T. Arkin I. Arkin I. T. Department of Biochemistry University of Cambridge 80 Tennis Court Road Cambridge CB2 1GA J.H. Bowie J.H. Bowie J.H. Bowie J. Bowie J. H. Department of Chemistry University of Adelaide North Terrace Adelaide, SA 5005 J. Torres J. Torres J. Torres J. Torres J. jtorres@ntu.edu.sg School of Biological Sciences Nanyang Technological University 1 Nanyang Walk 637616 Singapore J. Torres J. Torres Department of Chemistry University of Cambridge 80 Tennis Court Road Cambridge CB2 1GA jtorres@ntu.edu.sg Maculatin 1.1 is an antimicrobial peptide isolated from the Australian tree frog Litoria genimaculata that adopts an amphipathic, ?-helical structure in solution. Its orientation and conformation when incorporated to pre-formed DMPG (1,2-dimyristoyl-sn-glycero-3-phosphoglycerol) and DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) vesicles was determined using polarised Fourier transform infrared-attenuated total reflection infrared and deuterium exchange experiments. For DMPG membranes, our results show insertion of ?70% of the maculatin 1.1 molecules, with an angle of insertion of approximately 35° to the membrane normal and with a predominant ?-helical structure. These results suggest that maculatin 1.1 acts through a pore-forming mechanism to lyse bacterial membranes. A similar degree of insertion in DMPG (65%) and ?-helical structure was observed for a biologically inactive, less amphipathic maculatin 1.1 analogue, P15A, although the helix tilt was found to be greater (46°) than for maculatin 1.1. Similar experiments performed using DMPC liposomes showed poor insertion, less than 5%, for both maculatin 1.1 and its analogue. In addition, the shape of the amide I band in these samples is consistent with ?-helix, ?-structure and disordered structures being present in similar proportion. These results clearly show that maculatin 1.1 inserts preferentially in negatively charged membranes (DMPG) which mimic the negatively charged membrane of Gram-positive bacteria. We attribute the high percentage of insertion of the biologically inactive analogue in DMPG to the fact that its concentration on the membrane surface in our experiments is likely to be much higher than that found in physiological conditions. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved. FEBS Letters FEBS Lett. 00145793 FEBLA 2002 02 13 13 February 2002 1,2 dimyristoylglycero 3 phosphoglycerol antibiotic agent drug analysis drug comparison deuterium dimyristoylphosphatidylcholine glycerol derivative lipid liposome peptide drug analysis drug comparison protein derivative drug analysis drug comparison protein maculatin 1 drug analysis drug comparison protein P15A drug analysis drug comparison unclassified drug alpha helix article bacterial membrane channel gating concentration (parameters) drug activity electricity Fourier transformation Gram positive bacterium infrared radiation lipid membrane lysis physiology priority journal protein conformation protein structure structure analysis surface property Amino Acid Sequence Amphibian Proteins Anti-Bacterial Agents chemistry Antimicrobial Cationic Peptides chemistry Dimyristoylphosphatidylcholine Ion Channels chemistry Lipid Bilayers Models, Molecular Molecular Sequence Data Phosphatidylglycerols Protein Structure, Secondary Spectroscopy, Fourier Transform Infrared Anura Bacteria (microorganisms) Hylidae insertion sequences Litoria Litoria genimaculata Posibacteria 1303 1304 1312 99 37 4.2 4.7.3 Chiron Amphibian Proteins 0 Anti-Bacterial Agents 0 Antimicrobial Cationic Peptides 0 Dimyristoylphosphatidylcholine 13699-48-4 dimyristoylphosphatidylglycerol 61361-72-6 Ion Channels 0 Lipid Bilayers 0 maculatin-1.1 protein, Litoria 0 Phosphatidylglycerols 0 0032717048 R.M. Epand R.M. Epand H.J. Vogel H.J. Vogel Biochim. Biophys. Acta 0027328548 L.H. Lazarus L.H. Lazarus M. Atilla M. Atilla Prog. Neurobiol. 0023100945 M.G. Giovannini M.G. Giovannini L. Poulter L. Poulter B.W. Gibson B.W. Gibson D.H. Williams D.H. Williams Biochem. J. 0025860280 R.A. Cruciani R.A. Cruciani J.L. Baker J.L. Baker M. Zasloff M. Zasloff H.C. Chen H.C. Chen O.R.A. Colamonici O.R.A. Colamonici Proc. Natl. Acad. Sci. USA 0023875642 D. Marion D. Marion M. Zasloff M. Zasloff A. Bax A. Bax FEBS Lett. 0031062621 J. Gesell J. Gesell M. Zasloff M. Zasloff S.J. Opella S.J. Opella J. Biomol. NMR 0024438756 H. Duclohier H. Duclohier G. Molle G. Molle G. Spach G. Spach Biophys. J. 0002560639 H.V. Westerhoff H.V. Westerhoff D. Juretic D. Juretic R.W. Hendler R.W. Hendler M.H.V. Zasloff M.H.V. Zasloff Proc. Natl. Acad. Sci. USA 0025372569 D. Wade D. Wade A. Boman A. Boman B. Wahlin B. Wahlin C.M. Drain C.M. Drain D. Andreu D. Andreu H.G. Boman H.G. Boman R.B.D. Merrifield R.B.D. Merrifield Proc. Natl. Acad. Sci. USA 0032717886 Y. Shai Y. Shai Biochim. Biophys. Acta 0026969265 Y. Pouny Y. Pouny D. Rapaport D. Rapaport A. Mor A. Mor P. Nicolas P. Nicolas Y. Shai Y. Shai Biochemistry 0026742166 B. Bechinger B. Bechinger M. Zasloff M. Zasloff S.J. Opella S.J. Opella Biophys. J. 0027488740 B. Bechinger B. Bechinger M. Zasloff M. Zasloff S.J. Opella S.J. Opella Protein Sci. 0025993413 S. Frey S. Frey L.K. Tamm L.K. Tamm Biophys. J. 0032038181 T. Rozek T. Rozek R.J. Waugh R.J. Waugh S.T. Steinborner S.T. Steinborner J.H. Bowie J.H. Bowie M.J. Tyler M.J. Tyler J.J. Wallace J.J. Wallace Pept. Sci. 0034109713 B.C.S. Chia B.C.S. Chia J.A. Carver J.A. Carver T.D. Mulhern T.D. Mulhern J.H. Bowie J.H. Bowie Eur. J. Biochem. 0004003561 T.D. Brock T.D. Brock Biology of Microorganisms, 4th edn. Prentice-Hall, Englewood Cliffs, N.J. 0001160765 H. Lechevalier H. Lechevalier M.P. Lechevalier M.P. Lechevalier Microbiol. Lipids Ratledge, C. and Wilkinson, S.G., Eds, Academic Press, London 0026078046 R.C. MacDonald R.C. MacDonald R.I. MacDonald R.I. MacDonald B.M. Menco B.M. Menco K. Takeshita K. Takeshita N.K. Subbarao N.K. Subbarao L. Hu L. Hu Biochim. Biophys. Acta 0000337013 J.K. Kauppinen J.K. Kauppinen D.J. Moffat D.J. Moffat H.M. Mantsch H.M. Mantsch D.G. Cameron D.G. Cameron Appl. Spectrosc. 0029999396 I.T. Arkin I.T. Arkin W.P. Russ W.P. Russ M. Lebendiker M. Lebendiker S. Schuldiner S. Schuldiner Biochemistry 0003449930 N. Harrick N. Harrick Internal Reflection Spectroscopy Interscience Publishers, New York 0343092088 D. Marsh D. Marsh M. Muller M. Muller F.J. Schmitt F.J. Schmitt Biophys. J. 0001242101 M. Tsuboi M. Tsuboi J. Polym. Sci. 0032734050 D. Marsh D. Marsh Biophys. J. 0022691315 D.M. Byler D.M. Byler H. Susi H. Susi Biopolymers 0030819982 I.T. Arkin I.T. Arkin K.R. MacKenzie K.R. MacKenzie A.T. Brunger A.T. Brunger J. Am. Chem. Soc. 0026101392 C.E. Dempsey C.E. Dempsey R. Bazzo R. Bazzo T.S. Harvey T.S. Harvey I. Syperek I. Syperek G. Boheim G. Boheim I.D. Campbell I.D. Campbell FEBS Lett. 0026079128 C.E. Dempsey C.E. Dempsey B. Sternberg B. Sternberg Biochim. Biophys. Acta 0035954729 S. Fernandez-Lopez S. Fernandez-Lopez H.S. Kim H.S. Kim E.C. Choi E.C. Choi M. Delgado M. Delgado J.R. Granja J.R. Granja A. Khasanov A. Khasanov K. Kraehenbuehl K. Kraehenbuehl G. Long G. Long D.A. Weinberger D.A. Weinberger K.M. Wilcoxen K.M. Wilcoxen M.R. Ghadiri M.R. Ghadiri Nature