Bioactive Lipids

Also in the Oily Press Lipid Library:

Volume 16. Advances in Lipid Methodology – Five
Edited by Richard O. Adlof
Volume 15. Lipid Analysis (third edition)
Written by William W. Christie
Volume 14. Confectionery Fats Handbook
Written by Ralph E. Timms
Volume 13. Lipids for Functional Foods and Nutraceuticals
Edited by Frank D. Gunstone
Volume 12. Lipid Glossary 2
Written by Frank D. Gunstone and Bengt G. Herslöf
Volume 11. Lipids in Nutrition and Health: A Reappraisal
Written by Michael I. Gurr
Volume 10. Lipid Oxidation
Written by Edwin N. Frankel
Volume 9. Trans Fatty Acids in Human Nutrition
Edited by Jean Louis Sébédio and William W. Christie
Volume 8. Advances in Lipid Methodology – Four
Edited by William W. Christie
Volume 7. Advances in Lipid Methodology – Three
Edited by William W. Christie
Volume 6. Waxes: Chemistry, Molecular Biology and Functions
Edited by Richard J. Hamilton (out of print)
Volume 5. Lipids: Molecular Organization, Physical Functions and Technical
Applications
Written by Kåre Larsson
Volume 4. Advances in Lipid Methodology – Two
Edited by William W. Christie (out of print)
Volume 3. A Lipid Glossary (first edition)
Written by Frank D. Gunstone and Bengt G. Herslöf (out of print)
Volume 2. Advances in Lipid Methodology – One
Edited by William W. Christie (out of print)
Volume 1. Gas Chromatography and Lipids: A Practical Guide
Written by William W. Christie (out of print)
Bioactive Lipids
Edited by

ANNA NICOLAOU
School of Pharmacy
University of Bradford, UK
and

GEORGE KOKOTOS
Department of Chemistry
University of Athens, Greece

THE OILY PRESS

An imprint of PJ Barnes & Associates
Bridgwater, England

Published in association with

Lipid Technology
Published by The Oily Press
an imprint of PJ Barnes & Associates
PO Box 200, Bridgwater TA7 0YZ, England
Tel: +44-1823-698973, Fax: +44-1823-698971
E-mail: [email protected]
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ISBN 0-9531949-7-3

Copyright © 2004 PJ Barnes & Associates

All rights reserved. No part of this publication may be reproduced, stored in a retrieval
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All reasonable care is taken in the compilation of information for this book. However, the
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quential loss or loss of profits arising from the use of the information.

This book is Volume 17 in The Oily Press Lipid Library

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library

Typeset in 10½/12pt Times New Roman by

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Preface

There is no doubt that the elucidation of the human genome has changed the
face of modern science. All the resulting developments in gene analysis and
protein expression studies have emphasized the importance of a deeper
understanding of the regulatory mechanisms in the living cell and whole body.
Lipid research has not been left untouched by these advances. Intense research
effort has successfully defined the roles of lipids in cell signalling and the
pathophysiology of disease states. Lipidomics is emerging as a rapidly
expanding field aiming at the full characterization of lipid molecular species
and of their biological roles in terms of molecular mechanisms, gene
regulation, and protein structure and function.
For many people the term ‘lipid’ brings to mind negative associations
linked with excess dietary fat consumption, obesity and heart disease. How-
ever, lipids are essential components of the cell membrane shown to play
many dynamic roles in mediating and controlling a wide array of cellular
activities including membrane structure and organization, metabolic and
gene regulation, protein structure and function, energy production, and sig-
nalling pathways. Lipids have been intimately linked to the immune and
inflammatory responses, cell proliferation, mediation of programmed cell
death (apoptosis), as well as clearly shown to be major determinants in many
pathologies, including diabetes, cancer, cardiovascular disease and neuro-
degenerative disorders. Lipid metabolizing enzymes and rate limiting steps in
lipid-regulating and lipid-producing metabolic cascades have been targeted
for drug development, the best known example being the enzyme
cyclooxygenase, target for the widely used non-steroidal anti-inflammatory
drugs. Furthermore, functional foods and dietary supplements are already in
the market (e.g. fish oils, conjugated linoleic acid) making claims for health
benefits.
The main objective of this book is to present a clear overview of bioactive
lipids to scientists and technologists not totally familiar with lipid research.
We want to address the needs of new postgraduate researchers, industrial
technologists, physicians, and scientists in the pharmaceutical and food
industries. Overall, we wish to introduce this exciting field of research to all
who need to appreciate the multifaceted roles of these biomolecules in health
and disease. We have not tried to compile a series of scientific reviews but
have strived for chapters discussing the nomenclature, structures,
biochemistry, pharmacology and recent developments in the main classes of
bioactive lipids. Our starting point has been the definition of bioactive lipids
v
vi PREFACE

as lipids derived from components of the cellular membrane that mediate

cellular function.
The first chapter of the book deals with the properties and activities of fatty
acids. This is followed by detailed discussions of diacylglycerols and
phosphoinositides. The lysolipids are represented by sphingosine 1-phosphate
and lysophosphatidic acid, followed by an overview of the properties of ether
lipids. Ceramides and glycosphingolipids are covered in two chapters. The
metabolites of the arachidonic acid cascade are discussed in a further two
chapters on prostanoids, leukotrienes and lipoxins, followed by the last two
chapters of the book which focus on endocannabinoids and isoprostanes. The
authors come from several European countries and North America, each one
of them bringing on board their individual research and teaching experience,
and insight of their expertise.
The book would not have been possible without the help and patience of our
publisher, Peter J. Barnes, whom we thank. We are also thankful to William
Christie for helpful discussions and advice during the first stages of drafting
this book, the late David Horrobin for his suggestions and recommendations,
and, last but not least, our copy editor Beverley White who managed to convert
a bundle of manuscripts into a slick volume. We hope that this book will
contribute to the advancement of lipid research, will tempt more colleagues to
consider the roles of lipids in their individual fields, and inspire more students
to consider researching this fascinating area.

A. Nicolaou
G. Kokotos
May 2004
Contents

Preface v

List of Contributors xv

Glossary xvii

1 Fatty acids 1
PHILIP C. CALDER AND GRAHAM C. BURDGE
A. Introduction 1
B. Fatty acids – structure and nomenclature 2
C. Fatty acid biosynthesis 4
1. Biosynthesis of saturated fatty acids
2. Biosynthesis of monounsaturated fatty acids
3. Biosynthesis of polyunsaturated fatty acids
4. Biosynthesis of conjugated linoleic acid (CLA)
D. Lipid digestion, absorption and transport in the bloodstream and 7
delivery to tissues
1. Digestion and absorption of dietary lipids
2. Transport of fatty acids in the bloodstream and their delivery to tissues
E. The roles of fatty acids 12
1. Fatty acids as fuels
2. Fatty acids as membrane components
3. Fatty acylation of proteins
4. Fatty acids as eicosanoid precursors
5. Fatty acids and cell signaling
6. Fatty acids and gene expression
F. Fatty acids – nutritional aspects 21
1. Fatty acid composition of edible fats and oils and of foodstuffs
2. Intakes of fatty acids in humans
G. Fatty acids and human health 25
1. General comments
2. Essential fatty acid deficiency
3. The influence of dietary fatty acids on blood lipid and lipoprotein
concentrations

vii
viii CONTENTS

4. Long-chain n-3 PUFA and human health

5. A special requirement for DHA in brain development and function
6. Conjugated linoleic acid and human health
H. Concluding remarks 31
References 32

2 Diacylglycerols 37
KEVIN P. BECKER AND YUSUF A. HANNUN
A. Introduction 37
B. Glycerolipids 37
C. Formation of diacylglycerol during intermediary lipid metabolism 39
D. Regulated diacylglycerol metabolism 40
E. Diacylglycerols 41
1. Diacylglycerol derived from the hydrolysis of phosphoinositides
by phosphoinositide-specific phospholipase C
2. Diacylglycerol derived from the hydrolysis of phosphatidylcholine
through the action of phosphatidylcholine-specific
phospholipase D
3. Diacylglycerol derived from phosphatidylcholine through the
actions of phosphatidylcholine-specific phospholipase C
4. Production of diacylglycerol through sphingomyelin synthesis
F. Cellular targets of diacylglycerol 46
1. Protein kinase C isoenzymes
2. Non-kinase phorbol ester binding proteins
G. Role of diacylglycerol in normal cellular physiology 48
H. Role of diacylglycerol in pathophysiology 49
1. Role of diacylglycerol in tumor progression and cancer
2. Role of diacylglycerol in diabetes
3. Role of diacylglycerol in neurological diseases
4. Role of diacylglycerol in cardiac disease
5. Role of diacylglycerol and PKC in lymphoid function
I. Tools for analysis 52
1. Green fluorescent fusion proteins as probes for the targets of
action of cellular lipids
2. Lipid–protein interactions with fluorescence resonance energy
transfer
3. Diacylglycerol kinase assay/mass spectrometry
4. RNA interference for diacylglycerol metabolizing enzymes
5. Animal models
J. Monoacylglycerols 54
K. Alkylacylglycerols 55
L. Unanswered questions 55
 CONTENTS ix

1. When is diacylglycerol a metabolic intermediate versus a signaling

molecule?
2. What is the biological significance of non-kinase DAG/phorbol
ester receptors?
M. Concluding remarks 56
Acknowledgements 56
References 56

3 Phosphoinositides 63
BERNARD PAYRASTRE
A. Introduction 63
B. Biochemistry 63
1. The canonical pathway
2. The D-3 phosphorylated phosphoinositides
3. Interconversion between the polyphosphoinositides
4. Quantification of intracellular phosphoinositides
C. Localization of the various phosphoinositides 71
D. Functions of phosphoinositides 73
1. Phosphatidylinositol monophosphates: PtdIns(3)P, PtdIns(4)P and
PtdIns(5)P
2. Phosphatidylinositol bisphosphates: PtdIns(3,5)P2, PtdIns(3,4)P2
and PtdIns(4,5)P2
3. Phosphatidylinositol trisphosphate: PtdIns(3,4,5)P3
E. Phosphoinositides and human diseases 77
1. Phosphoinositide-dependent signalling and cancer
2. PtdIns(3,4,5)P3 5-phosphatase SHIP2 and type 2 diabetes
3. Myotubularin family and genetic diseases
4. The PtdIns(4,5)P2 5-phosphatase OCRL1 and Lowe syndrome
5. Phosphoinositides and pathogenic bacterial invasion
F. Concluding remarks 80
References 80

4 Lysolipids: Sphingosine 1-phosphate and lysophosphatidic acid 85

SUSAN PYNE
A. Introduction 85
B. Sphingosine 1-phosphate (S1P) 86
1. Metabolism of S1P
2. Intracellular actions of S1P
3. G-protein coupled receptors for S1P
C. Lysophosphatidic acid (LPA) 91
1. Metabolism of LPA
x CONTENTS

2. G-protein coupled receptors for LPA

D. Signalling of S1P and LPA 95
1. Signalling downstream of S1P and LPA receptors
2. Crosstalk and signal integration involving S1P and LPA
3. Termination of S1P/LPA signalling
E. Recent advances in therapeutics 98
F. Concluding remarks 100
References 100

5 Plasmalogens, platelet-activating factor, and other ether 107

glycerophospholipids
AKHLAQ A. FAROOQUI AND LLOYD A. HORROCKS
A. Introduction 107
B. Plasmalogens 109
1. Biosynthesis
2. Receptor-mediated degradation of plasmalogens
3. Roles of plasmalogens in mammalian metabolism
C. Platelet-activating factor (PAF) 115
1. Introduction
2. Biosynthesis
3. Degradation
4. Functions
D. Anti-tumor ether lipids 119
E. Other ether lipids 120
F. Involvement of plasmalogens, platelet-activating factor, and other 121
ether lipids in diseases
G. Concluding remarks 125
Acknowledgment 126
References 126

6 Ceramides 135
SILVIA VAENA DE AVALOS, JEFFREY A. JONES AND YUSUF A. HANNUN
A. Introduction 135
B. Metabolism 136
1. Nomenclature and structures of common sphingolipids
2. Specific pathways of sphingolipid metabolism
3. Comparison of sphingolipid metabolism between mammalian and
yeast cells
C. Experimental approaches and tools to probe for intracellular roles 142
of ceramide
1. Determination of endogenous levels of ceramide
 CONTENTS xi

2. Addition of exogenous ceramides to cells

3. Application of exogenous sphingomyelinases
4. Fusion proteins and targeting
5. Pharmacological manipulation of sphingolipid metabolism
6. Antibodies for the detection of enzymes of sphingolipid
metabolism
7. Antibodies directed against ceramide
8. Cloned enzymes
9. Knockout mice
10. Yeast as a model
11. RNA interference
D. Molecular mechanisms of ceramide action 149
E. Recent advances in understanding ceramide-regulated responses 153
1. Apoptosis
2. Oxidative stress
3. Heat stress
4. Ceramide and disease
F. Concluding remarks 159
Acknowledgements 159
References 160

7 Glycosphingolipids 169
THOMAS KOLTER
A. Introduction 169
B. Nomenclature and structures 170
1. Neutral and positively charged glycosphingolipids
2. Acidic glycosphingolipids
3. Nomenclature
4. Analysis and structure elucidation
C. Chemical synthesis of glycosphingolipids and analogues 173
D. Function 175
1. Structural diversity
2. Functional analysis
E. Biosynthesis 178
1. Sphingolipid biosynthesis
2. Biosynthesis of complex gangliosides
3. Genetically engineered mice
F. Degradation 183
1. Endocytosis
2. Activator proteins
3. Enzymology
4. Molecular mechanism of membrane digestion
xii CONTENTS

5. Glycosphingolipid storage diseases

G. Concluding remarks 191
References 192

8 Prostanoids 197
ANNA NICOLAOU
A. Introduction 197
B. Nomenclature and structures of prostanoids 198
C. Biosynthesis of prostanoids 200
1. Substrate availability: phospholipases A2
2. Cyclooxygenases (prostaglandin H synthases)
3. Prostanoid synthases
4. Cyclopentanone prostaglandins
5. Prostanoid catabolizing enzymes
D. Mechanisms of prostanoid action 207
1. Prostanoid transporters
2. Prostanoid receptors
3. Prostanoids and peroxisome proliferator-activated receptors
E. Biological activity and therapeutic applications 211
1. The role of prostanoids in inflammation
2. Prostanoids and the vascular system
3. Prostanoids and renal function
4. Prostanoids and the gastrointestinal system
5. Prostanoids and the reproductive system
6. The role of prostanoids in cancer
7. Novel nonsteroidal anti-inflammatory drugs (NSAID)
8. Prostanoids as therapeutic agents
F. Analytical methodologies for prostanoid detection 217
G. Concluding remarks 218
References 218

9 Leukotrienes and lipoxins 223

STEFANO FIORE
A. Introduction 223
B. Biosynthetic pathways 224
C. The leukotrienes 226
1. Leukotriene A4 (LTA4)
2. Leukotriene B4 (LTB4)
3. Peptido-leukotrienes
D. Lipoxins 229
E. Biological interfaces 231
 CONTENTS xiii

F. Determination of leukotrienes and lipoxins in biological samples 234

G. Concluding remarks 236
References 236

10 Endocannabinoids 245
GEORGE KOKOTOS 245
A. Introduction 245
B. Milestones in cannabinoid research 245
C. Cannabinoid receptors 246
D. Endocannabinoids and related fatty acid amides 248
E. Biosynthesis and metabolism 250
1. Biosynthesis
2. Fatty acid amide hydrolase
3. Monoglyceride lipase
4. Endocannabinoid transport
F. The endocannabinoid system as a target for the development of novel 255
therapeutic agents
1. Endocannabinoid analogues acting as cannabinoid receptor agonists
2. Fatty acid amide hydrolase inhibitors
3. Inhibitors of anandamide transport
G. Concluding remarks 259
References 259

11 Isoprostanes 265
SAMAR BASU
A. Introduction 265
B. Biosynthesis and occurrence 265
1. Biosynthesis
2. Occurrence
C. Metabolism and excretion 267
D. Isoprostanes as indicators of oxidant stress 271
E. Analytical methodologies 271
1. F2-isoprostane assays
2. Choice of biological fluid
3. Sample handling and storage
F. Pharmacological actions 273
G. Isoprostanes in human studies 274
1. Alcohol and smoking
2. Inflammation
3. Neurological disorders
4. Pulmonary diseases
xiv CONTENTS

5. Cardiovascular diseases
6. Diabetes
7. Reproductive disorders
8. Other studies
H. Isoprostanes as markers of lipid oxidation 276
I. Concluding remarks 278
References 278

Index 287
List of Contributors

Samar Basu, Associate Professor, Sections of Geriatrics & Clinical Nutrition

Research, Faculty of Medicine, Uppsala University, Box 609, SE-751 25
Uppsala, Sweden

Kevin P. Becker, Department of Biochemistry and Molecular Biology, Medical

University of South Carolina, 173 Ashley Avenue, PO Box 250509, Charleston,
SC 29425, USA

Graham Burdge, Senior Research Fellow, Institute of Human Nutrition,

School of Medicine, University of Southampton, Bassett Crescent East, South-
ampton SO16 7PX, UK

Philip C. Calder, Professor of Nutritional Immunology, Institute of Human

Nutrition, School of Medicine, University of Southampton, Bassett Crescent
East, Southampton SO16 7PX, UK

Akhlaq A. Farooqui, Research Scientist, Department of Molecular and Cellular

Biochemistry, The Ohio State University, 1645 Neil Avenue, Columbus,
OH 43210-1218, USA

Stefano Fiore, MD, University of Illinois at Chicago, Department of Medicine,

Section of Rheumatology, M/C 733, 900 S. Ashland Avenue, Chicago,
IL 60607-7171, USA

Yusuf A. Hannun, Professor and Chair, Department of Biochemistry and

Molecular Biology, Medical University of South Carolina, 173 Ashley Ave-
nue, PO Box 250509, Charleston, SC 29425, USA

Lloyd A. Horrocks, Professor Emeritus, Department of Molecular and Cellu-

lar Biochemistry, The Ohio State University, 1645 Neil Avenue, Columbus,
OH 43210-1218, USA

Jeffrey A. Jones, Research Associate, Department of Biochemistry and

Molecular Biology, Medical University of South Carolina, 173 Ashley Ave-
nue, PO Box 250509, Charleston, SC 29425, USA
xv
xvi CONTRIBUTORS

George Kokotos, Professor of Organic Chemistry, Laboratory of Organic

Chemistry, Department of Chemistry, University of Athens, Panepistimio-
polis 15771, Athens, Greece

Thomas Kolter, PhD, Kekulé-Institut für Organische Chemie und Biochemie

der Universität Bonn, Gerhard-Domagk-strasse 1, 53121 Bonn, Germany

Anna Nicolaou, Senior Lecturer in Pharmaceutical Chemistry, School of

Pharmacy, University of Bradford, Richmond Road, Bradford, West York-
shire BD7 1DP, UK

Bernard Payrastre, Director of Research at CNRS, Unité INSERM 563,

CPTP, Département d’Oncogenèse et Signalisation, Bâtiment B, CHU Purpan,
BP3028, 31024 Toulouse Cedex 3, France

Susan Pyne, Reader, Department of Physiology and Pharmacology, Strath-

clyde Institute for Biomedical Sciences, University of Strathclyde, 27 Taylor
Street, Glasgow G4 ONR, UK

Silvia Vaena de Avalos, PhD, Department of Biochemistry and Molecular

Biology, Medical University of South Carolina, 173 Ashley Avenue, PO
Box 250509, Charleston, SC 29425, USA