M e t h o d s i n M o l e c u l a r B i o l o g y ™

Series EditorJohn M. Walker

School of Life SciencesUniversity of Hertfordshire

Hatfield, Hertfordshire, AL10 9AB, UK

For other titles published in this series, go to www.springer.com/series/7651

Polyamines

Methods and Protocols

Edited by

Anthony E. Pegg

Department of Cellular and Molecular Physiology, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA, USA

Robert A. Casero, Jr.

The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA

EditorsAnthony E. Pegg, Ph.D.College of MedicineMilton S. Hershey Medical CenterPennsylvania State UniversityUniversity Drive 500Hershey, PA 17033-0850, USAapegg@psu.edu

Robert A. Casero, Jr., Ph.D.School of MedicineSidney Kimmel Comprehensive Cancer Center Johns Hopkins UniversityOrleans St. 1650Baltimore, MD 21231, USArcasero@jhmi.edu

ISSN 1064-3745 e-ISSN 1940-6029ISBN 978-1-61779-033-1 e-ISBN 978-1-61779-034-8DOI 10.1007/978-1-61779-034-8Springer New York Dordrecht Heidelberg London

Library of Congress Control Number: 2011921314

© Springer Science+Business Media, LLC 2011All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden.The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights.

Printed on acid-free paper

Humana Press is part of Springer Science+Business Media (www.springer.com)

v

Preface

Polyamines are ubiquitous cellular components that perform multiple functions and are essential for normal growth and development. Polyamine biosynthesis and the regulation of polyamine levels, which are closely linked to cell growth, have been the subject of many studies over the past 40 years. A volume of this series (Vol. 79, Polyamine Protocols) was published in 1998 and described some methods for assays of biosynthetic and catabolic enzymes, measurement of polyamine levels, and transport measurements. In the last decade, there have been important new findings in the polyamine field and a variety of new experi-mental systems have become available. These include studies with animal models and human patients, which indicate that polyamines play a critical role in normal development and in the development of neoplasia. Therapeutic roles for polyamine inhibitors and ana-logs have been established and more are under investigation. Polyamine metabolism is now recognized as a significant source of oxidative damage. Polyamines are now established as regulators of critical ion channels. Polyamine transport systems have been characterized much more fully. The polyamine family has been expanded to include compounds present in plants, thermophilic microorganisms, and protozoal parasites that play essential roles in their physiology. Methods for identifying polyamine-responsive genes are now available. The proposed volume will cover methodologies for studies in these areas.

Anthony E. Pegg, PhDRobert A. Casero, Jr., PhD

vii

Contents

Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vContributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

Part I IntroductIon

1 Current Status of the Polyamine Research Field . . . . . . . . . . . . . . . . . . . . . . . . . . 3Anthony E. Pegg and Robert A. Casero, Jr.

Part II IdentIfIcatIon of PolyamIne metabolIc enzymes and characterIzatIon of PolyamIne-regulated genes and Ion channels

2 Exploring Polyamine Biosynthetic Diversity Through Comparative and Functional Genomics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39Anthony J. Michael

3 Characterization of Genes for Polyamine Modulon . . . . . . . . . . . . . . . . . . . . . . . . 51Kazuei Igarashi and Keiko Kashiwagi

4 Posttranscriptional Regulation of Gene Expression in Epithelial Cells by Polyamines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67Lan Xiao and Jian-Ying Wang

5 Identification, Chemical Synthesis, and Biological Functions of Unusual Polyamines Produced by Extreme Thermophiles . . . . . . . . . . . . . . . . . 81Tairo Oshima, Toshiyuki Moriya, and Yusuke Terui

6 Polyamine Block of Inwardly Rectifying Potassium Channels . . . . . . . . . . . . . . . . 113Harley T. Kurata, Wayland W.L. Cheng, and Colin G. Nichols

Part III transgenIc models for study of PolyamIne functIon

7 Carcinogenesis Studies in Mice with Genetically Engineered Alterations in Polyamine Metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 129David J. Feith

8 Transgenic Rodents with Altered SSAT Expression as Models of Pancreatitis and Altered Glucose and Lipid Metabolism . . . . . . . . . . . . . . . . . . 143Anne Uimari, Mervi T. Hyvönen, Eija Pirinen, and Leena Alhonen

9 Use of (Gyro) Gy and Spermine Synthase Transgenic Mice to Study Functions of Spermine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159Xiaojing Wang and Anthony E. Pegg

viii Contents

Part IV enzymes InVolVed In PolyamIne catabolIsm and PosttranslatIonal ProteIn modIfIcatIon

10 A Simple Assay for Mammalian Spermine Oxidase: A Polyamine Catabolic Enzyme Implicated in Drug Response and Disease . . . . . . . . . . . . . . . . 173Andrew C. Goodwin, Tracy R. Murray-Stewart, and Robert A. Casero, Jr.

11 Characterization, Assay, and Substrate Specificity of Plant Polyamine Oxidases . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183Panagiotis N. Moschou and Kalliopi A. Roubelakis-Angelakis

12 Assay of Deoxyhypusine Synthase Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195Edith C. Wolff, Seung Bum Lee, and Myung Hee Park

13 Assay of Deoxyhypusine Hydroxylase Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207Jong Hwan Park, Edith C. Wolff, and Myung Hee Park

Part V regulatIon of PolyamIne content

14 Identification and Assay of Allosteric Regulators of S-Adenosylmethionine Decarboxylase. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 219Erin K. Willert, Lisa N. Kinch, and Margaret A. Phillips

15 Protocols for Studying Antizyme Expression and Function . . . . . . . . . . . . . . . . . . 237Noriyuki Murai, Yasuko Murakami, and Senya Matsufuji

16 Identification, Assay, and Functional Analysis of the Antizyme Inhibitor Family . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 269Chaim Kahana

17 Posttranscriptional Regulation of Ornithine Decarboxylase . . . . . . . . . . . . . . . . . . 279Shannon L. Nowotarski, Sofia Origanti, and Lisa M. Shantz

Part VI PolyamIne transPort and uPtake

18 Identification and Assays of Polyamine Transport Systems in Escherichia coli and Saccharomyces cerevisiae . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295Keiko Kashiwagi and Kazuei Igarashi

19 Genetic and Biochemical Analysis of Protozoal Polyamine Transporters . . . . . . . . 309Marie-Pierre Hasne and Buddy Ullman

20 Heparan Sulfate Proteoglycan-Mediated Polyamine Uptake . . . . . . . . . . . . . . . . . 327Johanna Welch, Katrin Svensson, Paulina Kucharzewska, and Mattias Belting

21 Polyamine Transport Systems in Mammalian Cells and Tissues . . . . . . . . . . . . . . . 339Takeshi Uemura and Eugene W. Gerner

22 Procedures to Evaluate the Importance of Dietary Polyamines . . . . . . . . . . . . . . . 349Paul Acheampong, Mary J. Macleod, and Heather M. Wallace

Part VII PolyamInes and dIsease

23 Determination of N1,N12-Diacetylspermine in Urine: A Novel Tumor Marker . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367Masao Kawakita, Kyoko Hiramatsu, Mari Yanagiya, Yosuke Doi, and Mieko Kosaka

ixContents

24 Spermidine/Spermine-N1-Acetyltransferase in Kidney Ischemia Reperfusion Injury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 379Kamyar Zahedi and Manoocher Soleimani

25 Use of Polyamine Metabolites as Markers for Stroke and Renal Failure . . . . . . . . . 395Kazuei Igarashi and Keiko Kashiwagi

26 Methods to Evaluate Alterations in Polyamine Metabolism Caused by Helicobacter pylori Infection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 409Alain P. Gobert, Rupesh Chaturvedi, and Keith T. Wilson

27 High-Resolution Capillary Gas Chromatography in Combination with Mass Spectrometry for Quantification of Three Major Polyamines in Postmortem Brain Cortex. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 427Gary Gang Chen, Laura M. Fiori, Orval A. Mamer, and Gustavo Turecki

28 Spermine Synthase Deficiency Resulting in X-Linked Intellectual Disability (Snyder–Robinson Syndrome) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 437Charles E. Schwartz, Xaiojing Wang, Roger E. Stevenson, and Anthony E. Pegg

Part VIII chemIstry and analysIs

29 Methylated Polyamines as Research Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 449Alex R. Khomutov, Janne Weisell, Maxim A. Khomutov, Nikolay A. Grigorenko, Alina R. Simonian, Merja R. Häkkinen, Tuomo A. Keinänen, Mervi T. Hyvönen, Leena Alhonen, Sergey N. Kochetkov, and Jouko Vepsäläinen

30 Fluorescent Substrates for Polyamine Catabolic Enzymes and Transport . . . . . . . . 463Koichi Takao and Akira Shirahata

31 Use of Polyamine Derivatives as Selective Histone Deacetylase Inhibitors . . . . . . . 475Patrick M. Woster

32 Measurement of Polyamine pKa Values . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 493Ian S. Blagbrough, Abdelkader A. Metwally, and Andrew J. Geall

33 Polyamine Analysis by LC-MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 505Merja R. Häkkinen

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 519

xi

Contributors

Paul acheamPong • Division of Applied Medicine, University of Aberdeen, Aberdeen, Scotland, UK

leena alhonen • A .I . Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Eastern finland, Kuopio, Finland

mattIas beltIng • Department of Clinical Sciences, Section of Oncology, Lund University and Lund University Hospital, Lund, Sweden

Ian s. blagbrough • Department of Pharmacy and Pharmacology, University of Bath, Bath, UK

robert a. casero, Jr. • School of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA

ruPesh chaturVedI • Division of Gastroenterology, Hepatology, and Nutrition, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA

gary gang chen • McGill Group for Suicide Studies, Douglas Mental Health University Institute, McGill University, Montreal, QC, Canada

Wayland W.l. cheng • Department of Cell Biology and Physiology, Washington University School of Medicine, St . Louis, MO, USA

yosuke doI • Alfresa Pharma Corporation, Osaka, JapandaVId J. feIth • Department of Cellular and Molecular Physiology,

Pennsylvania State University College of Medicine, Hershey, PA, USAlaura m. fIorI • McGill Group for Suicide Studies, Douglas Mental Health

University Institute, McGill University, Montreal, QC, CanadaandreW J. geall • Department of Pharmacy and Pharmacology, University of Bath,

Bath, UKeugene W. gerner • Arizona Cancer Center, University of Arizona,

Tucson, AZ, USAalaIn P. gobert • UR454 Unite de Microbiologie, INRA,

Saint-Genes-Champanelle, France andreW c. goodWIn • The Sidney Kimmel Comprehensive Cancer Center,

Johns Hopkins University School of Medicine, Baltimore, MD, USAnIkolay a. grIgorenko • BASF, GVP/SI, Basel, SwitzerlandmerJa r. häkkInen • Department of Biosciences, Laboratory of Chemistry,

Biocenter Kuopio, University of Kuopio, Kuopio, FinlandmarIe-PIerre hasne • Department of Biochemistry and Molecular Biology,

Oregon Health and Science University, Portland, OR, USAkyoko hIramatsu • Department of Molecular Medical Research,

Tokyo Metropolitan Institute of Medical Science, Tokyo, JapanmerVI t. hyVönen • Department of Biotechnology and Molecular Medicine,

A .I . Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Kuopio, Kuopio, Finland

xii Contributors

kazueI IgarashI • Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan; Amine Pharma Research Institute, Innovation Plaza at Chiba University, Chiba, Japan

chaIm kahana • Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot, Israel

keIko kashIWagI • Faculty of Pharmacy, Chiba Institute of Science, Chiba, Japanmasao kaWakIta • Department of Molecular Medical Research, Tokyo Metropolitan

Institute of Medical Science, Tokyo, Japantuomo a. keInänen • Department of Biotechnology and Molecular Medicine,

A .I . Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Kuopio, Kuopio, Finland

lIsa n. kInch • Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA

alex r. khomutoV • Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991, Russia

maxIm a. khomutoV • Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

sergey n. kochetkoV • Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

mIeko kosaka • Alfresa Pharma Corporation, Osaka, JapanPaulIna kucharzeWska • Department of Clinical Sciences, Section of Oncology,

Lund University and Lund University Hospital, Lund, Swedenharley t. kurata • Department of Anesthesiology, Pharmacology, and Therapeutics,

University of British Columbia, Vancouver, BC, Canadaseung bum lee • The Oral and Pharyngeal Cancer Branch, NIDCR,

National Institutes of Health, Bethesda, MD, USAmary J. macleod • Division of Applied Medicine, University of Aberdeen,

Aberdeen, Scotland, UKorVal a. mamer • Mass Spectrometry Unit, McGill University, Montreal,

QC, Canadasenya matsufuJI • Department of Molecular Biology, The Jikei University

School of Medicine, Tokyo, Japanabdelkader a. metWally • Department of Pharmacy and Pharmacology,

University of Bath, Bath, UKanthony J. mIchael • University of Texas Southwestern Medical Center,

Department of Pharmacology, Forest Park, Dallas, TX, USAtoshIyukI morIya • Institute of Environmental Microbiology, Kyowa-kako Co .,

Machida, Tokyo, Japan; Department of Molecular Biology, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo, Japan

PanagIotIs n. moschou • Department of Biology, University of Crete, Heraklion, Greece

norIyukI muraI • Department of Molecular Biology, The Jikei University School of Medicine, Tokyo, Japan

yasuko murakamI • Department of Molecular Biology, The Jikei University School of Medicine, Tokyo, Japan

xiiiContributors

tracy r. murray-steWart • The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, USA

colIn g. nIchols • Department of Cell Biology and Physiology, Washington University School of Medicine, St . Louis, MO, USA

shannon l. noWotarskI • Department of Cellular and Molecular Physiology, Milton S . Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA, USA

sofIa orIgantI • Department of Cellular and Molecular Physiology, Milton S . Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA, USA

taIro oshIma • Institute of Environmental Microbiology, Kyowa-kako Co ., Machida, Tokyo, Japan

Jong hWan Park • The Oral and Pharyngeal Cancer Branch, NIDCR, National Institutes of Health, Bethesda, MD, USA

myung hee Park • The Oral and Pharyngeal Cancer Branch, NIDCR, National Institutes of Health, Bethesda, MD, USA

anthony e. Pegg • College of Medicine, Milton S . Hershey Medical Center, Pennsylvania State University, Hershey, PA, USA

margaret a. PhIllIPs • Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA

eIJa PIrInen • A .I . Virtanen Institute for Molecular Sciences, Biocenter Kuopio, University of Kuopio, Kuopio, Finland

kallIoPI a. roubelakIs-angelakIs • Department of Biology, University of Crete, Heraklion, Greece

charles e. schWartz • J .C . Self Research Institute, Greenwood Genetic Center, Greenwood, SC, USA

lIsa m. shantz • Department of Cellular and Molecular Physiology, Milton S . Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA, USA

akIra shIrahata • Department of Clinical Dietetics and Human Nutrition, Laboratory of Cellular Physiology, Josai University, Saitama, Japan

alIna r. sImonIan • Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia

manoocher soleImanI • Department of Surgery, Division of Nephrology and Hypertension, University of Cincinnati College of Medicine, Cincinnati, OH, USA

roger e. steVenson • J .C . Self Research Institute, Greenwood Genetic Center, Green-wood, SC, USA

katrIn sVensson • Department of Clinical Sciences, Section of Oncology, Lund University and Lund University Hospital, Lund, Sweden

koIchI takao • Laboratory of Cellular Physiology, Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmaceutical Sciences, Josai University, Saitama, Japan

yusuke teruI • Faculty of Pharmacy, Chiba Institute of Science, Choshi, Chiba, Japan

xiv Contributors

gustaVo tureckI • Douglas Mental Health University Institute, McGill University, Montreal, QC, Canada

takeshI uemura • Arizona Cancer Center, University of Arizona, Tucson, AZ, USAanne uImarI • A .I . Virtanen Institute for Molecular Sciences, Biocenter Kuopio,

University of Kuopio, Kuopio, Finlandbuddy ullman • Department of Biochemistry and Molecular Biology, Oregon

Health and Science University, Portland, OR, USAJouko VePsäläInen • Department of Biosciences, Biocenter Kuopio, University

of Kuopio, Kuopio, Finlandheather m. Wallace • Division of Applied Medicine, University of Aberdeen,

Aberdeen, Scotland, UKJIan-yIng Wang • Baltimore Veterans Affairs Medical Center, Baltimore, MD, USAxIaoJIng Wang • Department of Cellular and Molecular Physiology,

Milton S . Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey, PA, USA

Janne WeIsell • Department of Biosciences, Biocenter Kuopio, University of Kuopio, Kuopio, Finland

Johanna Welch • Department of Clinical Sciences, Section of Oncology, Lund University and Lund University Hospital, Lund, Sweden

erIn k. WIllert • Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX, USA

keIth t. WIlson • Division of Gastroenterology, Hepatology, and Nutrition, Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA; Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN, USA

edIth c. Wolff • The Oral and Pharyngeal Cancer Branch, NIDCR, National Institutes of Health, Bethesda, MD, USA

PatrIck m. Woster • Department of Pharmaceutical Sciences, Wayne State University, Detroit, MI, USA

lan xIao • Baltimore Veterans Affairs Medical Center, Baltimore, MD, USAmarI yanagIya • Alfresa Pharma Corporation, Osaka, Japankamyar zahedI • Department of Surgery, Division of Nephrology and Hypertension,

University of Cincinnati College of Medicine, Cincinnati, OH, USA