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Subcellular Biochemistry Volume 18 Intracellular Parasites
SUBCELLULAR BIOCHEMISTRY
SERIES EDITOR J. R. HARRIS, Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Martinsried bei Miinchen, Germany
ASSISTANT EDITOR H. J. HILDERSON, University of Antwerp, Antwerp, Belgium
Recent Volumes in This Series:
Volumes 5-11 Edited by Donald B. Roodyn
Volume 12
Volume 13
VQ1ume 14
Volume 15
Volume 16
Volume 17
Volume 18
Volume 19
Immunological Aspects Edited by J. R. Harris
Fluorescence Studies on Biological Membranes Edited by H. J. HUderson
Artificial and Reconstituted Membrane Systems Edited by J. R. Harris and A.-H. Etemadi
Virally Infected Cells Edited by J. R. Harris
Intracellular Transfer of Lipid Molecules Edited by H. J. HUderson
Plant Genetic Engineering Edited by B. B. Biswas and J. R. Harris
Intracellular Parasites Edited by J. L. Avila and J. R. Harris
Endocytic Components: Identification and Characterization Edited by J. J. M. Bergeron and J. R. Harris
A Continuation Order Plan is available for this series. A continuation order will bring delivery of each new volume immediately upon publication. Volumes are billed only upon actual shipment. For further information please contact the publisher.
Subcellular Biochemistry Volume 18 Intracellular Parasites
Edited by
J. L. Avlla Instituto de Biomedicina Caracas, Venezuela
and J. R. Harris Max-Planck-Institute for Biochemistry Martinsried bei Miinchen, Germany
Springer Science+Business Media, LLC
The Library of Congress cataloged the first volume of this title as follows:
Sub-cellular biochemistry.
London, New York, Plenum Press. v. illus. 23 cm. quarterly.
Began with Sept. 1971 issue. Cf. New serial titles. 1. Cytochemistry—Periodicals. 2. Cell organelles—Periodicals.
QH611.S84 574.8/76 73-643479
ISBN 978-1-4899-1653-2 ISBN 978-1-4899-1651-8 (eBook) DOI 10.1007/978-1-4899-1651-8
This series is a continuation of the journal Sub-Cellular Biochemistry, Volumes 1 to 4 of which were published quarterly from 1972 to 1975
© 1992 Springer Science+Business Media New York Qrginally published by Plenum Press, New York in 1992 Softcover reprint of the hardcover 1st edition 1992
All rights reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher
INTERNATIONAL ADVISORY EDITORIAL BOARD
J. L. AVILA, Instituto de Biomedicina, Caracas, Venezuela
J. J. M. BERGERON, McGill University, Montreal, Canada
B. B. BISWAS, Bose Institute, Calcutta, India
N. BORGESE, CNR Center for Pharmacological Study, Milan, Italy
M. J. COSTELLO, University of North Carolina, Chapel Hill, North Carolina, USA
N . CRAWFORD, Royal College of Surgeons, London, England
A.-H. ETEMADI, University of Paris VI, Paris, France
W. H. EVANS, National Institute for Medical Research, London, England
H. GLAUMANN, Karolinska Institute, Huddinge, Sweden
D. R. HEADON, University College Galway, Galway, Ireland
P. L. J0RGENSEN, University of Aarhus, Aarhus, Denmark
J. KIM, Osaka University, Osaka, Japan
J. B. LLOYD, University of Keele, Keele, England
J. A. LUCY, Royal Free Hospital School of Medicine, London, England
A. H. MADDY, University of Edinburgh, Edinburgh, Scotland
J. D. MORRE, Purdue University, West Lafayette, Indiana, USA
P. QUINN, King's College London, London, England G. RALSTON, The University of Sydney, Sydney, Australia
S. ROTTEM, The Hebrew University, Jerusalem, Israel
M. R. J. SALTON, New York University Medical Center, New York, New York, USA G. SCHATTEN, University of Wisconsin-Madison, Madison, Wisconsin, USA F. S. SJOSTRAND, University of California-Los Angeles, Los Angeles, California, USA
F. WUNDERLICH, University of Diisseldorf, Diisseldorf, Germany
G. ZAMPIGHI, University of California-Los Angeles, Los Angeles, California, USA
I. B. ZBARSKY, Russian Academy of Sciences, Moscow, Russia
Contributors
Marie Laure Ancelin Montpellier, France
Membrane Interactions, CNRS URA 530, 34095
Jose Luis Avila Venezuela
Instituto of Biomedicina, Apartado 4043, Caracas 10 lOA ,
Patrick Bastien Laboratoire d'Ecologie Medicale et Pathologie Parasitaire, Annex de la Faculte de Medecine, 34000 Montpellier, France
Christine Blaineau Laboratoire d'Ecologie Medicale et Pathologie Para-sitaire, Annex de la Faculte de Medecine, 34000 Montpellier, France
Clement Bordier Switzerland
Affinity Products, CH-I023 Crissier-sur-Lausanne,
Juan Jose Cazzulo Instituto de Investigaciones Bioquimicas "Luis F. Leloir," Fundaci6n Campomar-CONICET-Facultad de Ciencias Exaetas y Naturales, Universidad de Buenos Aires, 1405 Buenos Aires, Argentina
Robert Etges Institute of Biochemistry, University of Lausanne, CH-I066 Epalinges, Switzerland
Brian Fenton Cell Cycle Genetics Group, Department of Biochemistry, Medical Sciences Institute, University of Dundee, Dundee DDI 4HN, Scotland
Laurie Hall Department of Tropical Public Health, Harvard School of Pub-lic Health, Boston, Massachusetts 02115
Selma M. B. Jeronimo Department of Biochemistry, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte 59072, Brazil
Mark A. Jutila Veterinary Molecular Biology, Marsh Laboratory, Montana State University, Bozeman, Montana 59717
vii
viii Contributors
Paul B. Kimsey Division of Comparative Medicine, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
J. Joseph Marr Discovery Research, Searle Pharmaceuticals, Skokie, llli-nois 60077, and Department of Internal Medicine, Rush Medical College, Chicago, lllinois 60612
Mary McGurn Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115
Steven R. Meshnick Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, Michigan 48109
Michel Pages Centre de Recherche de Biochimie Macromoleculaire du CNRS et Unite 249 de I'INSERM, 34033 Montpellier Cedex, France
Richard D. Pearson Division of Geographic Medicine, Departments of Internal Medicine and Pathology, University of Virginia School of Medicine, Charlottesville, Virginia 22908
Laura Povinelli Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115
Linda D. Saffer Department of Pathology, University of Virginia School of Medicine, Charlottesville, Virginia 22908
Pascal Schneider Institute of Biochemistry, University of Lausanne, CH-I066 Epalinges, Switzerland
Joseph D. Schwartzman Department of Pathology, University of Virginia School of Medicine, Charlottesville, Virginia 22908
Anuraj Shankar Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115
Alwi M. Shatry Veterinary Molecular Biology, Marsh Laboratory, Montana State University, Bozeman, Montana 59717
Clarence A. Speer Veterinary Molecular Biology, Marsh Laboratory, Mon-tana State University, Bozeman, Montana 59717
Cynthia M. Theodos Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115
Richard G. Titus Department of Tropical Public Health, Harvard School of Public Health, Boston, Massachusetts 02115
Salvatore J. Turco Department of Biochemistry, University of Kentucky Medical Center, Lexington, Kentucky 40536
Contributors ix
Henri J. Vial France
Membrane Interactions, CNRS URA 530, 34095 Montpellier,
David WaUiker Institute of Cell, Animal, and Population Biology, Univer-sity of Edinburgh, Edinburgh EH9 3JN, Scotland
Earl Weidner Department of Zoology and Physiology, Louisiana State Uni-versity, Baton Rouge, Louisiana 70803, and The Marine Biological Laboratory, Woods Hole, Massachusetts 02543
Michael W. White Veterinary Molecular Biology, Marsh Laboratory, Mon-tana State University, Bozeman, Montana 59717
Preface
Volume 18 of the Subcellular Biochemistry series, which specializes in various aspects of the biochemistry of the intracellular parasites, was initially proposed by Jose Luis Avila and strongly supported by myself, as Series Editor of Subcellular Biochemistry. Considerable assistance was received from Professor Frank Wunderlich (University of Dusseldorf) and more particularly from Dr. Michael Miles (London School of Hygiene and Tropical Medicine) during the compilation of the list of possible chapters. Our initial aim was to present a comprehensive survey of this broad field of study. Although some interesting topics have been lost due to authors backing out late in the production schedule of the book, the manuscripts that were submitted have provided a useful overview of the subject, with notable strength within the field of Leishmania. The 13 chapters of the book have been grouped according to subject. The first five chapters deal with Leishmania and are followed by two chapters on Tryponosoma cruzi, two on the malarial parasites, and two on the Coccidia. The fmal two chapters cover the Microsporidia and chemotherapy, respectively.
The book opens with the contribution of Selma Jeronimo and Richard Pearson, in which they set the scene by discussing the Leishmania within the context of adaption to extracellular and intracellular survival. Chapter 2 is a detailed account of the membrane proteins and enzymes of Leishmania by Pascal Schneider, Clement Bordier, and Robert Etges. In Chapter 3, Salvatore Turco presents an account of structural and functional aspects of the lipophosphoglycan of Leishmania. Immunity to leishmaniasis and the role of T cells and cytokines are dealt with in an extremely competent manner by Richard Titus and his colleagues. The fmal chapter on the Leishmania comes from Patrick Bastien, Christine Blaineau, and Michel Pages, who discuss at some length the important topic of molecular karyotype analysis. Moving to Tryponosoma cruzi, Jose Luis Avila presents a thorough account of the role of intracellular digestion of endocytosed proteins as a source of amino acids for protein synthesis, and Juan Jose Cazzulo discusses energy metabolism. A lengthy overview of the malarial lipids comes from Henri Vial and Marie Laure Ancelin and is followed by the contribution of Brian Fenton and David Walliker on the genetic analysis of malaria
xi
xii Preface
parasites. The first of the two chapters on the coccidian parasites, from Joseph Schwartzman and Linda Saffer, considers the mechanisms of Toxoplasma gondii entry and exit from host cells. Then Michael White and his colleagues present a survey of both biochemical and ultrastructural observations on coccidian host cell interactions. Earl Weidner then covers the microsporidian parasites, with emphasis on the cytoskeletal proteins in the Thelohania species and Spraguea lophii. In the final chapter of the book, Steven Meshnick and Joseph Marr appropriately present a survey of new developments in chemotherapy; they deal successively with drugs directed at the four main groups of intracellular parasites discussed in the earlier chapters: malaria, leishmaniasis, American trypanosomiasis, and toxoplasmosis.
Thus, despite the unforeseen omissions mentioned above, this volume nevertheless possesses a considerable unity of purpose and content. It is hoped that the book will be of interest and use to all those involved with the investigation of intracellular parasites and treatment of the diseases they cause in humans. Biochemists interested in the cellular metabolism and membrane systems of intracellular parasites should also find much of interest in this book, as will molecular geneticists.
Robin Harris Martinsried bei Miinchen. Germany
Contents
Chapter 1 The Leishmania: Protozoans Adapted for ExtraceUular and IntraceUular Survival
Selma M. B. Jeronimo and Richard D. Pearson
1. Introduction.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 2. Taxonomy of Leishmania Species ........................... 2 3. Clinical Leishmaniasis .................................... 4
3.1. Visceral Leishmaniasis .............................. 5 3.2. Cutaneous Leishmaniasis ............................ 7 3.3. Mucosal Leishmaniasis .............................. 7
4. Life Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 4.1. Amastigotes....................................... 8 4.2. Promastigotes...................................... 8
5. Metabolism.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 5.1. Carbohydrates..................................... 12 5.2. Amino Acids ...................................... 14 5.3. Glyoxylate Cycle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 5.4. Fatty Acids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
6. Nucleotide Metabolism ................................... 15 6.1. Purines........................................... 15 6.2. Pyrimidines....................................... 17
7. Proton Pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 8. Heat Shock Proteins. . . .. . . . . .. . ... . . . . . .. . .. . .. . . . . . . . . .. 18 9. Surface Molecules ....................................... 19
9.1. Glycoprotein 63 kDa . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
xiii
~ Con~nu
9.2. Phosphomonoesterases .............................. 20 9.3. ATPases.......................................... 21 9.4. Protein Kinases .................................... 21 9.5. Lipophosphoglycan................................. 22
10. Leishmania-Macrophage Interactions. . ... .. . . . . ... . . ... . . . . . 22 10.1. Attachment........................................ 22 10.2. Ingestion of Promastigotes ........................... 25 10.3. Intracellular Survival ................................ 25
11. Immunology of Leishmaniasis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 12. References.............................................. 29
Chapter 2 Membrane Proteins and Enzymes of LeishmIJnia
Pascal Schneider, Clement Bordier, and Robert Etges
1. Introduction...... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 1.1. Biology of Leishmania .............................. 39 1.2. Organelles of Leishmania. . . .... . . . . . ..... . . . .. . ..... 41 1.3. Membrane Proteins of Leishmania. . ..... .. . . . ... ... .. . 42 1.4. Metabolite Uptake and Energy Generation .............. 43
2. Proteins and Enzymes of the Plasma Membrane ............... 45 2.1. ATPases.......................................... 45 2.2. Symports and Antiports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 2.3. Uniports.......................................... 48 2.4. Plasma Membrane Enzyme Activities .................. 49
3. Membrane Proteins of Intracellular Organelles ................ 55 3.1. Mitochondrial Proton Pumps ......................... 56 3.2. Mitochondrial FoP. ATP Synthase. . .... . . ... . ... . .. . . . 56 3.3. Mitochondrial Ca2 +-Transporter and ER Ca2 +-ATPase .... 56 3.4. Olycosomal Integral Membrane Proteins. . ... . .. . . ... .. . 57
4. Membrane Protein Adaptations ............................. 57 4.1. The Promastigote ....... . . . . . . ..... . . .. . ... . . ... .. . . 57 4.2. The Amastigote .................................... 60
5. Membrane Proteins as Therapeutic Thrgets in Leishmaniasis ..... 60 5.1. Chemotherapy..................................... 60 5.2. Vaccine Development ............................... 61 5.3. Diagnostic Tools ................................... 63
6. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 7. References.................. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Contents
Chapter 3 The Lipophosphoglycan of Leishmania
Salvatore 1. Thrco
xv
1. Introduction.............. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73 2. Structure of LPG ........................................ 74 3. Cellular Aspects ......................................... 78 4. Developmental Modification of LPG during Metacyclogenesis .. . . 79 5. Participation of LPG in Parasite-Macrophage Surface
Interactions ................... ; . . . . . . . . . . . . . . . . . . . . . . . . . 82 6. Intracellular Functions of LPG ............................. 83
6.1. Requirement of LPG for Intracellular Survival ........... 83 6.2. Effects of LPG on Action of Hydrolases ................ 83 6.3. Effects of LPG on the Oxidative Burst ................. 85
7. Immunological Implications of LPG . . . . . . . . . . . . . . . . . . . . . . . . . 87 8. Other Unusual Parasitic Glycoconjugates ..................... 89
8.1. LPG-like Glycoconjugates in Leishmania ............... 89 8.2. Other Unusual Lipid-Containing Glycoconjugates in
Parasites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91 9. Future Considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
10. References........ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92
Chapter 4 Role of T Cells in Immunity to the Intracellular Pathogen, Leishmania major Richard G. Titus, Cynthia M. Theodos, Paul B. Kimsey, Anuraj Shankar, Laurie Hall, Mary McGurn, and Laura Povinelli
1. General Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99 2. T Cells and Leishmaniasis ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103
2.1. Early Work. . . . . . ... . .. . . .. . . . . . .. . . ... . . . ...... . . . 103 2.2. CD4+, CD8+, and 'Y8-TCR+ T Cells in Leishmaniasis... 107
3. Cytokines and Leishmaniasis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111 3.1. Introduction....................................... 111 3.2. Role of Interferon-'Y, Interleukin-2, and Interleukin-4 ..... 112 3.3. Role of Thmor Necrosis Factor. . . . . . . . . . . . . . . . . . . . . . . . 116 3.4. Role of Other Cytokines ............................. 118
4. Vaccination.......... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120 5. Conclusions............. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 6. References.............................................. 123
Chapter 5 Molecular Karyotype Analysis in Leishmania Patrick Bastien, Christine Blaineau, and Michel Pages
Contents
1. Introduction................ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131 1.1. The Parasite....................................... 131 1.2. Pulsed Field Electrophoresis. . .... . . . . . . . ..... . ... . . . . 133 1.3. Molecular Karyotype and Chromosomes ................ 134 1.4. Aims and Scope of This Chapter ...................... 135
2. The Molecular Karyotype of Leishmania ..................... 136 2.1. The Complete Karyotype of Two Cloned Strains of
Leishmania. . . ...... . . . . . . .... . . . . . . . .... . . ... . . . . . 136 2.2. Stability of the Molecular Karyotype ................... 140 2.3. The Karyotype of a Leishmania Strain Is Representative
of One Clone. . .. . . .. . . . . . . . .... . . . . . . . .... . .. . . . . . 141 2.4. Genome Size ...................................... 141 2.5. Recognition of the Different Chromosomes and
Chromosomal Homologies ........................... 142 3. Molecular Karyotype Polymorphisms in Leishmania. . . ... .... . . 146
3.1. Polymorphisms between Different Species .............. 147 3.2. Polymorphisms within the Same Species. . . . ..... . .. . . . . 147 3.3. Interclonal (Intrastrain) Polymorphisms ................. 153 3.4. Chromosome Size Polymorphisms and Size-Variable
Homologues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154 3.5. PFE and the Taxonomy of Leishmania ................. 158 3.6. Relative Homogeneity of the Karyotype in Leishmania .... 162
4. Molecular Basis for the Karyotype Polymorphisms ............. 164 4.1. Intra- Rather Than Interchromosomal Rearrangements ..... 164 4.2. DNA Amplifications and Deletions .................... 165 4.3. Recurrent Chromosomal Rearrangements. . ... . .... . . .. .. 166 4.4. Subtelomeric Location of Rearrangements . . . . . . . . . . . . . . . 167 4.5. Interchromosomal Exchange. . . . . . ... . . ... . . .... . . .... 169 4.6. De Novo Appearance of Small Chromosomes............ 170
5. Interpretation at the "Population" Level ..................... 171 6. Implications and Outlook . . . . . .. .. .. . . .. . . . .. . . . . .. ... . . . .. 173
6.1. Functional Significance of the Karyotype Polymorphisms .. 173 6.2. Implications on the Ploidy of Leishmania ............... 174 6.3. On the Presence of Genetic Exchange in Leishmania.. . . . . 176
7. Summary and Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 178 8. References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 179
Contents
Chapter 6 Intracellular Digestion of Endocytosed Proteins as a Source of Amino Acids for Protein Synthesis in Trypanosoma cruzi Jose Luis Avila
xvii
1. Introduction and Background. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189 2. Trypanosoma cruzi . . . .. . .. . . .... . . . . .... ... .. .. . . . . . .. . . . 190
2.1. Biology of Trypanosoma cruzi ........................ 190 2.2. Protein and Amino Acid Requirements in
Trypanosoma cruzi ................................. 192 3. The Endocytic System of Trypanosoma cruzi ................. 195
3.1. Morphological Aspects .............................. 195 3.2. Dynamic Aspects of Endocytosis in Trypanosoma cruzi ... 202
4. Pinocytosis in Trypanosoma cruzi .. . . . . . . . . . . . . . . . . . . . . . . . . . 203 4.1. Time Course and Temperature-Dependent Bovine Liver
Catalase Binding to Trypanosoma cruzi Epimastigotes and Trypomastigotes ................................ 203
4.2. Concentration-Dependent Acquisition of [125I]-BLC .. . .. . . 204 4.3. Uptake of FITC-Dextran and Lucifer Yellow by
Trypanosoma cruzi Epimastigotes and Trypomastigotes .... 205 4.4. Influence of Bovine Liver Catalase on the Uptake of
Other Proteins ..................................... 205 4.5. Bovine Liver Catalase Uptake Rate Values Obtained in
T. cruzi Epimastigotes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 205 4.6. Effect of Amino Acids and ATP on FITC-Dextran
Accumulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207 4.7. Uptake of the Lysosomotropic Drug N-Dodecyl Imidazol by
T. cruzi Epimastigotes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210 5. Intracellular Degradation of Proteins in Trypanosoma cruzi ...... 210
5.1. Proteases Present in Trypanosoma cruzi Epimastigotes .... 210 5.2. Evidence of Lysosomal Digestion of Endocytosed Proteins
by Trypanosoma cruzi Epimastigotes ................... 213 5.3. Effect of Basic Substances, Ionophores, and Proteolytic
Inhibitors on Trypanosoma cruzi Epimastigote Multiplication ..................................... 213
5.4. Influence of pH on the in Vitro Hydrolysis of Bovine Liver Catalase and Bovine Serum Albumin by Trypanosoma cruzi Epimastigote Homogenates ........................... 217
6. Evidence of Lysosome-Released Amino Acids as Building Blocks for Protein Synthesis in Trypanosoma cruzi Epimastigotes ....... 219 6.1. Release of Lysine from Exogenous Protein Sources ....... 219
xviii Contents
6.2. Influence of Exogenous Amino Acid Levels on the Lysine Release from Endogenous Protein Sources .............. 220
6.3. Influence of Exogenous Amino Acid Levels on the Lysine Release from Exogenous Protein Sources ............... 221
7. Intracellular Protein Degradation in Trypanosoma cruzi Epimastigotes ........................................... 224 7.1. Enhanced Intracellular Proteolysis Provides Amino Acids for
de Novo Protein Synthesis ........................... 224 8. Concluding Remarks ..................................... 228 9. References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
Chapter 7 Energy Metabolism in Trypanoso1lUl cruz;
Juan Jose Cazzulo
1. Introduction..... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 235 2. Transport and Utilization of Carbohydrate, Proteins, and
Amino Acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 236 3. Carbohydrate Catabolism: The Aerobic Fermentation of
Glucose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 237 3.1. End Products of Glucose Catabolism ................... 237 3.2. Enzyme Pathways for Glucose Catabolism .............. 238 3.3. Systems for Reoxidation of Glycolytic NADH ........... 240 3.4. Subcellular Compartmentation and Regulation of
Glucose Catabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 244 4. Oxidative Phosphorylation ................ " .. . . ... . . . .. . . . 247
4.1. Phosphorylation in Vivo and in Submitochondrial Particles .......................................... 247
4.2. Mitochondrial ATPase. . . . . . . . . . . . . . . . . . . .. . . . .. . . . . . 247 5. Protein and Amino Acid Catabolism. . . . . . . . . . . . .. . . . .. . . . .. . 248
5.1. Production of NH3 . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . 248 5.2. Proteolytic Enzymes ................................ 248 5.3. Amino Acid Catabolism ............................. 249
6. Relationship between Carbohydrate and Amino Acid Catabolism ............................................. 251
7. References..................................... . . . . . . . . . 252
Chapter 8 Malarial Lipids: An Overview
Henri J. Vial and Marie Laure Ancelin
1. 2.
Lipids: What For? Structure of Lipids
259 261
Con~nu ~
3. Lipids of Plasmodium and the Surrounding Plasma. .. . . . . .. . . . . 263 3.1. Plasma Lipids during Malarial Infection ................ 264 3.2. What Is the Role of Cholesterol? ...................... 265 3.3. Neutral Lipids and PL in Infected Erythrocytes .......... 267 3.4. Glycolipids in Infected Erythrocytes ................... 268 3.5. Lipids of Various Fractions of the Infected Erythrocyte .... 269
4. Biosynthetic Metabolic Pathways of Glycerolipids ............. 271 4.1. Fatty Acid Sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 273 4.2. Biosynthesis of PC ................................. 275 4.3. Biosynthesis of PE ................................. 277 4.4. Biosynthesis of PS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 278 4.5. Biosynthesis of PI .................................. 278 4.6. Can These Metabolic Pathways Provide All the PL Needed
by the Parasite? .................................... 279 4.7. Equilibrium between the Different Pathways for PL
Biosyntheses and PL Compartmentalization ............. 281 4.8. Adjustment of the Lipid Composition .................. 283 4.9. Conclusion........................................ 284
5. Phospholipid Metabolism: An Original Target for New Malaria Chemotherapy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
6. Structural Organization of Lipids in the Host Cell Membrane .... 290 6.1. Functional Modifications of the Host Cell Membrane May
Involve Changes in Lipids ........................... 290 6.2. Asymmetric Distribution of the PL .................... 291 6.3. Transbilayer Mobility of PL .......................... 295
7. Dynamics of Lipids within Infected Cells .................... 295 8. Summary and Perspectives ................................ 297 9. References.............................................. 299
Chapter 9 Genetic Analysis of Malaria Parasites
Brian Fenton and David Walliker
1. Introduction.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 307 2. Life Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 308
2.1. The Malaria Life Cycle and Its Relationship to Genetic Studies .................................... 308
3. Genetic Analysis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 310 3.1. Introduction....................................... 310 3.2. Inheritance of Specific Characters ..................... 311
4. Conclusion........... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 324 5. References.............................................. 326
Contents
Chapter 10 How Toxoplasma gondii Gets In and Out of Host Cells
Joseph D. Schwartzman and Linda D. Saffer
1. Toxoplasma gondii: The Universal Coccidian ................. 333 1.1. Importance of Toxoplasmosis ......................... 333 1.2. The Process of Host Cell Invasion ..................... 334 1.3. Active Invasion versus Phagocytosis ................... 335 1.4. The Parasitophorous Vacuole ......................... 335
2. Morphology of Invasion and Egress ......................... 337 2.1. Light Microscopy .................................. 337 2.2. Ultrastructure...................................... 343
3. Motility of T. gondii ..................................... 343 3.1. Cytoskeleton of Coccidians . . . . . . . . . . . . . . . . . . . . . . . . . . . 347 3.2. Possible Motors and Mechanisms ............... '" . . . . 349
4. Factors Secreted at the Time of Invasion ..................... 350 4.1. Penetration-Enhancing Factor ......................... 350 4.2. Apical Organelles .................................. 350 4.3. Unlocalized Factors...... . . . ..... . . . . . . . . . .... . ..... 356
5. Conclusions............................................. 358 6. References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
Chapter 11 Biochemical and Ultrastructural Observations of Coccidian Parasite and Host CeU Interactions Michael W. White, Alwi M. Shatry, Mark A. Jutila, and Clarence A. Speer
1. Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365 2. Coccidian Biology ....................................... 366 3. Ultrastructural Alterations in Infected Host Cells . . . . . . . . . . . . . . . 369 4. Initial Parasite and Host Cell Interactions . . . . . . . . . . . . . . . . . . . . . 372
4.1. Adherence and Penetration ........................... 372 4.2. Influence of Parasites on Endocytosis .................. 375
5. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 380 6. References.............................................. 381
Chapter 12 Cytoskeletal Proteins Expressed by Microsporidian Parasites
Earl Weidner
1. Introduction....... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 385 1.1. Microsporidians as Intracellular Parasites ............... 385
Contents xxi
1.2. Microsporidians in Epithelial Cells .................... 385 1.3. Microsporidian Range in Phyletic Host Groups .......... 386 1.4. Microsporidians with Sporophorous Vesicles. . . . . . . . . . . . . 386 1.5. Cytoskeletal Proteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 386
2. Cytoskeletal Proteins of Vertebrate Keratinocytes and Thelohania Spore Sacs ............................................. 386 2.1. Cytokeratins, Desmosomal Proteins, and Involucrin . . . . . . . 386 2.2. Thelohania Species Cytokeratin and Desmosomal Protein
Expression ........................................ 388 2.3. The Cytoskeletal Analogue Structure in TheLohania
Species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 389 2.4. Origins of Desmoplakin-Cytokeratin Assemblages in
TheLohania Species ................................. 390 3. Intermediate Filament Cage Assembly in Spraguea Lophii ....... 391
3.1. Intermediate Filament Cage Assemblages ............... 391 3.2. The Cytoskeletal Envelope on S. Lophii Spores. . . . ... . . .. 393 3.3. Cytokeratins, Clathrin, and Calmodulin in the
S. Lophii Cage ..................................... 395 3.4. Discussion of S. Lophii IF Cage Assemblage. . . . . . ... .... 395 3.5. Inactivation of the S. Lophii Spore Extrusion Apparatus. . . . 397
4. References........ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 397
Chapter 13 IntraceUular Parasites: New Developments in Chemotherapy
Steven R. Meshnick and J. Joseph Marr
1. Introduction........... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 1 2. Malaria................................................ 401
2.1. Background....................................... 401 2.2. Biochemistry of Plasmodia. .... . . . ... ... . . . ..... . . . . . 403 2.3. Quinoline Methanols ................................ 404 2.4. 4-Aminoquinolines ................................. 406 2.5. 8-Aminoquinolines ................................. 408 2.6. Antifolates: Sulfa Drugs ............................. 409 2.7. Antifolates: Dihydrofolate Reductase Inhibitors .......... 411 2.8. Artemisinin (Qinghaosu) and Derivatives ............... 412 2.9. Miscellaneous Agents ............................... 414
3. Leishmaniasis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 416 3.1. Background....................................... 416 3.2. Microbiology...................................... 417 3.3. Pathogenesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 417 3.4. Current Treatment .................................. 418
Contents
3.5. Experimental Chemotherapy. ...... . . . . .... . .... . ..... 419 4. American Trypanosomiasis (Chagas' Disease) ................. 425
4.1. Background....................................... 425 4.2. Compounds of Current Clinical and Experimental Interest. . 425
5. Toxoplasmosis........................................... 432 5.1. Background....................................... 432 5.2. Microbiology...................................... 433 5.3. Treatment......................................... 433
6. References.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 434
Index..................................................... 443
