Sphingolipid Biochemistry Handbook of Lipid Research Editor: Donald J

Handbook of 3 Lipid Research

Sphingolipid Biochemistry Handbook of Lipid Research Editor: Donald j. Hanahan The University of Texas Health Center at San Antonio San Antonio, Texas

Volume 1 Fatty Acids and Glycerides Edited by Amis Kuksis

Volume 2 The Fat-Soluble Vitamins Edited by Hector F. DeLuca

Volume 3 Sphingolipid Biochemistry Julian N. Kanfer and Sen-itiroh Hakomori Handbook of 3 Lipid Research

Sphingolipid Biochemistry

Julian N. Kanfer University ojManitoba Faculty ojMedicine Winnipeg, Manitoba, Canada and Sen-itiroh Hakomori Fred Hutchinson Cancer Research Center and University oj Washington Seattle, Washington

Plenum Press · New HJrk and London ISBN 978-1-4757-0398-6 ISBN 978-1-4757-0396-2 (eBook) DOI 10.1007/978-1-4757-0396-2

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 Preface

Interest in and emphasis upon different aspects of the sphingolipids have, in general, followed the biochemical developments of the day. The early inves• tigators were preoccupied principally with the isolation of "pure" compounds and structural elucidation. This historical perspective is found in the discus• sion presented in Chapter 1 (Section 1.1.2 and Table III). Still, the isolation and structural characterization of glycolipids are the basic foundation of all our knowledge of enzymology, immunology, and cell biology. Recent infor• mation obtained on structure has greatly affected the interpretation of various phenomena related to glycolipids. New structures suggest a new role of gly• colipids as antigens and receptors. Ten years ago, only four neutral glycolipids and two gangliosides were known in human erythrocytes. We now know structures of at least twenty additional neutral glycolipids and ten additional gangliosides in human erythrocytes that are known to be important blood group, heterophil, and autoantigens. Erythrocytes are only one example of a cell type whose glycolipid profile has been extensively studied. Our defective knowledge in immunology and cell biology may be due to incomplete un• derstanding of structural chemistry. Modern methodology based on methyla• tion analysis, mass spectrometry, and enzymatic degradation has supple• mented classical analysis based on clorimetry. Nuclear magnetic resonance spectroscopy is still in the development stage, but will eventually replace var• ious chemical analyses. However, important future studies should be directed toward elucidating the organizational structure of glycolipids in membranes. Knowledge of the biosynthesis of the sphingolipids has also followed a classical pattern of development. Soon after the availability of radioisotopes, early in vivo studies provided insight into the pathways of biosynthesis. Perhaps the most significant contribution was by Saul Roseman and his colleagues who demonstrated the logical sequential addition of carbohydrates from their nu• cleotide derivatives to produce these glycosphingolipids in vitro. A number of laboratories subsequently participated in providing the details of the individ• ual reactions responsible for the production of the simplest compound, sphin• gosine, and eventually for that of the more complex gangliosides. Even though these pathways are accepted as dogma and appear in most general textbooks of biochemistry, the enzymes responsible for the individual reactions have not been purified. The data have been derived largely from studies employing particles, frequently of ill-defined character. Information about these reac• tions is presented in Chapter 2.

v vi Preface There is a wealth of information on the enzymatic degradation of the sphingolipids. Most of the enzymes catalyzing the hydrolysis of particular bonds have been purified, some to homogeneity. This contrasts with the lesser amount of information available on biosynthetic activity and can be readily ascribed to two circumstances: The hydrolytic enzymes do not appear to be hydrophobic proteins firmly embedded in a membrane matrix, and, therefore, are relatively simple to "solubilize," making them amenable to purification. A pioneer in providing descriptive information about the sphingolipid hydro• lases is Shimon Gatt. Secondly, the existence of a number of sphingolipid storage diseases, caused by diminished hydrolytic activity, has provided the required "relevancy" to enhance the likelihood of grant support. A discussion of the hydrolytic activity and the sphingolipidoses is presented in Chapters 2 and 3. The early workers could not foresee the impact of studies on the individual hydrolases which became an important contribution to, and essen• tial building block for, the fields of prenatal diagnosis, clinical genetics, and mass population screening, and which stimulated discussions on the floor of the United States Congress. Investigations into the nature of tissue antigens revealed that many were lipid-soluble molecules. Early workers realistically preferred working with aqueous soluble compounds and devoted their efforts to the isolation and elucidation of the various blood group substances. Greater progress on the structural basis of various glycolipid antigens has been achieved with the availability of reliable immunological methods that can be applied to glyco• lipids, and by various modern methods in structural analysis (see Chapter 1). A summary of our knowledge on the structural concept of glycolipid specificity is discussed in Chapter 4. Of particular interest is the introduction of the monoclonal antibody approach and the discovery of tumor-associated gly• colipid markers. We are at an early stage of development and Chapter 4 could only be an introduction to the coming great age of glycolipid immunology. The realization that glycolipids are present on the external surface of cell membranes and that they possess antigenic activity has brought them into the modern-day arena of cell biology. Here again, their potential relevance to human disease has resulted in great activity on the sphingolipids. There are numerous reports of modifications in glycolipid composition and metabolism associated with oncogenic transformation. A clear-cut dependence on differ• entiation and development has also been demonstrated in more recent studies. There are indications that glycolipid changes are caused by activation of trans• forming genes. Studies in this area have inspired hope that the role of gly• colipids in regulation of cell growth and cell recognition would eventually be realistic. Although our knowledge in this area is extremely premature and fragmentary, it is reviewed in Chapter 4. The external aspect of the plasma membrane is also believed to contain the complement of cellular receptors for a diversity of agents. Since this coincides with the location of the glycosphingolipids, there has been a flurry of activity in the attempt to correlate receptors and glycolipids. This is de• scribed in the last Chapter. Preface vii It is difficult to predict the probable future developments, but it seems reasonable to predict that a component of future efforts will be a continuation of present ones. However, revolutionary developments can be expected in the application of monoclonal antibodies, physical instrumentation analysis, and gene cloning. A precise analysis of glycolipid organization in membranes, roles in cell growth regulation, cell adhesion and cell recognition, and their genetic and epigenetic control will be exciting developments for the near future. The knowledge of glycolipid function will be extremely useful in prevention and treatment of various human diseases. There will probably not be any great effort expended on the biosynthesis of the sphingolipids, mainly because the individual steps have been accepted and are "ancient history." Some groups may think it important actually to purify and study the isolated enzymes responsible for the individual reactions. However, genetic control of glycosyltransferases at transcriptional and trans• lationallevels and epigenetic regulation of the assembly of glycosyltransferases in membranes will be the central theme of glycolipid studies in the future. Various diseases, including cancer, inflammation, and aging, may well be related to abnormalities of these processes. There will be continued isolation and structural elucidation of glycolipids. The work will be focused on many uncharacterized species which are available only in small quantities in specific types of cells or tissues and are limited to a specific stage of development. There has been a diminution of the previous flurry of activity on the sphingolipidoses largely because the enzyme deficiencies are understood, the patients can be identified, pedigrees can be screened, and reliable prenatal diagnosis can be offered. Perhaps the future efforts will reveal the patho• physiological basis for the characteristic disease process resulting from the accumulation of a particular sphingolipid. The techniques of recombinant DNA technology and monoclonal antibodies will undoubtably be utilized to reveal the primary genetic abnormalities responsible for the separate diseases. A good deal of current interest centers upon the anecdotal patient who pre• sents uniquely to the physician. These patients may provide useful materials and challenges to accepted dogma so that more incisive investigations can be carried out. The more difficult areas for prediction, but probably the most exciting, must be in cell biology. Perhaps some day we will be able to speak with some degree of certainty about the function of these intriguing molecules. This book was written to fill a need for a single source of information in this field. It was not written for the sphingolipid specialist, although it is hoped that even the specialist will find some useful information. It was prepared for those who are curious to learn about these intriguing molecules. Immunol• ogists, cell biologists, physicians, biochemists, educators, and students will find it useful for their various personal needs. Inevitably, a few of our colleagues may have some degree of dissatisfaction for a number of possible reasons. We may have missed a contribution, we may disagree with them, we may misinterpret them, we may be incorrect in viii Preface a statement, or our style may not coincide with theirs. Nonetheless, a text such as this inevitably reflects the authors' biases, experience, and particular research interests. We have attempted to be comprehensive, but not neces• sarily encyclopedic. We wish to thank the many researchers who have been actively engaged in the study of the sphingolipids for providing so much interesting data for us to read, understand, and digest. S. Hakomori is greatly indebted to Mrs. Charlotte Pagni for reference arrangement and typing of the manuscript, to Drs. Kiyohiro Watanabe, Wil• liam W. Young, and Reiji Kannagi for their collaboration in providing ref• erences and figures, and to Dr. Roger A. Laine for reading Chapter 1. Section 1.3.2 was based partially on a review written in Japanese by Drs. A. Hayashi and T. Matsubara. He is particularly grateful to a number of scientists and publishers who allowed him to cite published materials and to reproduce various figures.

Julian N. Kanfer Sen-itiroh Hakomori Contents

Chapter 1 Chemistry of Glycosphingolipids Sen-itiroh H akomori 1.1. Introduction ...... 1 1.1.1. Classification and Nomenclature of Glycosphingolipids .. 1 1.1.2. Brief History of the Chemistry of Glycosphingolipids .... 6 1.2. Isolation of Glycosphingolipids ...... 16 1.2.1. Extraction...... 16 1.2.2. Purification of Lipid Extract: Elimination of Nonlipid Contaminants ...... 18 1.2.3. Separation of Gangliosides and Long-Chain Neutral Glycolipids ...... 19 1.2.4. Separation of Individual Glycolipids ...... 22 1.3. Characterization of Glycosphingolipids ...... 32 1.3.1. Analysis by Thin-Layer Chromatography...... 33 1.3.2. Characterization of Ceramide Structure, Fatty Acids, and Long-Chain Bases...... 37 1.3.3. Determination of Carbohydrate Composition ...... 43 1.3.4. Release of Oligosaccharides from Glycosphingolipids .... 49 1.3.5. Determination of Carbohydrate Sequence ...... 51 1.3.6. Determination of the Position of Glycosyl Linkages ...... 57 1.3.7. Determination of Anomeric Configuration (a or 13) in Glycolipids by Chromium Trioxide Oxidation ...... 61 1.3.8. Direct-Probe Mass Spectrometry of Glycolipids ...... 62 1.3.9. Spectrometric Analysis of Glycosphingolipids: Infrared (IR), Nuclear Magnetic Resonance, and Electron Spin Resonance Spectra ...... 76 1.4. Structure of Glycosphingolipids ...... 89 1.4.1. Structural Variation in Ceramides ...... 90 1.4.2. Simpler Glycosphingolipids: Ceramide Monohexosides, Ceramide Dihexosides, and Sulfatides ...... 94 1.4.3. Globo-Series Glycolipids ...... 99 1.4.4. Lacto-Series Glycolipids ...... 101 1.4.5. Muco-Series Glycolipids: Glycolipids with Digalactosyl to Hexagalactosyl Core Structure ...... 115 1.4.6. Simpler Gangliosides and Hematosides: Sialosyl Glycolipids without Hexosamines ...... 117 Contents 1.4.7. Ganglio-Series Glycolipids: Gangliosides with Ganglio-N• triose, Ganglio-N-tetraose, and Ganglio-N-pentaose Structure ...... 121 1.4.8. Glycosphingolipids of Water-Living Invertebrates ...... 129 1.4.9. Plant Sphingolipids (Phytoglycosphingolipids) ...... 134 1.5. Chemical Synthesis and Modification of Sphingosines and Glycosphingolipids ...... 136 1.5.1. Synthesis of Long-Chain Bases ...... 138 1.5.2. Synthesis of Glycosphingolipids ...... 144 1.6. Pioneers in Glycolipid Chemistry ...... 145 1.7. References...... 150

Chapter 2 Sphingolipid Metabolism Julian N. Kanfer 2.1. Sphingosine Bases ...... 167 2.1.1. In Vivo Studies...... 167 2.1.2. In Vitro Studies ...... 168 2.1.3. In Vivo Studies on Sphingosine Base Utilization ...... 181 2.2. The Psychosines ...... 183 2.2.1. Galactosylsphingosine Formation ...... 184 2.2.2. Glucosylsphingosine Formation...... 186 2.2.3. Galactosylsphingosine Acylation ...... 186 2.2.4. Glucosylsphingosine Acylation...... 187 2.3. Ceramide (N-Acylsphingosine) ...... 187 2.4. Galactosylceramide ...... 194 2.4.1. In Vivo Studies ...... 194 2.4.2. Biosynthesis in Vitro ...... 195 2.4.3. Hydrolysis in Vitro ...... 198 2.5. Ceramide-galactoside-3-sulfate (Sulfatide) ...... 200 2.5.1. In Vivo Studies ...... 200 2.5.2. Biosynthesis in Vitro ...... 203 2.5.3. Hydrolysis in Vitro ...... 206 2.6. Glucosylceramide...... 207 2.6.1. Biosynthesis in Vitro ...... 208 2.6.2. Hydrolysis in Vitro ...... 209 2.7. Ganglioside Metabolism ...... 211 2.7.1. In Vivo Studies ...... 211 2.7.2. Biosynthesis in Vitro ...... 215 2.8. Sphingomyelin ...... 230 2.8.1. Biosynthesis in Vitro ...... 230 2.8.2. Hydrolysis in Vitro ...... 234 2.9. Biosynthesis of the Sphingolipids: Summary ...... 235 2.10. Hydrolysis of the Sphingolipids: Summary ...... 238 2.11. References ...... 240 Contents

Chapter 3 The Sphingolipidoses Julian N. Kanfer 3.1. Farber's Lipogranulomatosis ...... 252 3.1.1. Chemical Studies ...... 252 3.1.2. Enzyme Defect ...... 253 3.2. Krabbe's Disease (Globoid Cell Leukodystrophy) ...... 253 3.2.1. Clinical ...... 253 3.2.2. Pathology ...... 255 3.2.3. Biochemistry ...... 257 3.3. Metachromatic Leukodystrophy...... 262 3.3.1. Clinical ...... 262 3.3.2. Pathology ...... 264 3.3.3. Biochemistry ...... 264 3.4. Gaucher's Disease...... 267 3.4.1. Clinical ...... 268 3.4.2. Pathology ...... 269 3.4.3. Biochemistry ...... 270 3.5. Fabry's Disease (Angiokeratoma Corporis Diffusium) ...... 278 3.5.1. Clinical ...... 278 3.5.2. Pathology ...... 279 3.5.3. Biochemistry ...... 279 3.6. The Gangliosidoses ...... 283 3.6.1. GM 2 Gangliosidosis Type I (Classical Tay-Sachs' Disease) 283 3.6.2. GM 2 Gangliosidosis Type II (Sandhoff-Jatzkewitz Variant) 284 3.6.3. GM 2 Gangliosidosis Type III Quvenile) ...... 284 3.6.4. GM 1 Gangliosidosis Type I (Pseudo-Huder's, Landing's Syndrome; Neurovisual Lipidosis) ...... 296 3.6.5. GM 1 Gangliosidosis Type II Quvenile GM 1 Gangliosidosis; Derry's Syndrome) ...... 297 3.7. Niemann-Pick's Disease ...... 301 3.7.1. Clinical ...... 301 3.7.2. Pathology ...... 302 3.7.3. Biochemistry ...... 302 3.8. GM3 Gangliosidosis ...... 309 3.8.1. Pathology ...... 310 3.8.2. Biochemistry ...... 310 3.9. General Comments ...... 310 3.10. References ...... 312

Chapter 4 Glycosphingolipids in Cellular Interaction, Differentiation, and Oncogenesis Sen-itiroh Hakomori 4.1. Introduction ...... 327 4.2. Organization and Dynamic State of Glycolipids in Membranes .. 328 xii Contents 4.2.1. Crypticity and Organization of Glycolipids ...... 328 4.2.2. Association of Membrane Proteins and Glycolipids ...... 329 4.2.3. Dynamic Behavior of Glycolipids in Membranes ...... 334 4.2.4. Common Carbohydrate Chain in Glycolipids and Glycoproteins ...... 336 4.3. Glycolipids in Cellular Interaction and Differentiation ...... 337 4.3.1. Cellular Interaction ...... 337 4.3.2. Glycolipids as Differentiation Markers ...... 342 4.4. Role of Glycolipids in Cell Growth Control ...... 349 4.4.1. Cell Contact Response of Glycolipids as Related to "Contact Inhibition" ...... 349 4.4.2. Cell Cycle and Mitogenesis ...... 353 4.4.3. Glycolipid Addition in Cell Culture ...... 353 4.4.4. Modification by Antiglycolipid Antibodies ...... 355 4.4.5. Glycolipid Changes Caused by Differentiation Inducers 355 4.5. Glycolipid Changes in Oncogenic Transformation: Deficiency in Glycolipid Function ...... 356 4.5.1. Common Features of Glycolipid Changes ...... 360 4.5.2. Studies with Temperature-Sensitive Mutants...... 361 4.5.3. Comparison of Tumor Cells with Different Degrees of Tumorigenicity, Malignancy, and Metastatic Capability .. 361 4.5.4. Chemical Carcinogenesis and Glycolipids ...... 365 4.5.5. Studies of Human Cancer ...... 366 4.5.6. Search for Glycolipid Tumor Antigens or Cell-Surface Markers ...... 367 4.5.7. Biological Significance of Tumor-Associated Glycolipid Changes ...... 368 4.6. Glycolipids as Possible Mediators of Immune Response ...... 370 4.6.1 Modulation of B-Cell or T-Cell Response by Gangliosides 370 4.6.2. Gangliosides as Mediators of Immune Cell Recognition.. 371 4.6.3. Glycolipids as Lymphokine Receptors ...... 371 4.7. Postscript: EnIgmas Concerning Glycolipid Functions ...... 372 4.8. References...... 373

Chapter 5 Glycolipid Antigens and Genetic Markers Sen-itiroh Hakomori and William W. Young, Jr. 5.1. Introduction ...... 381 5.2. General Properties of Glycolipid Antigens ...... 383 5.2.1. Unique Immunogenicity and Antigenicity of Glycolipids ...... 383 5.2.2. Immune Response to Glycolipid Antigens ...... 386 5.2.3. Methods for Detecting Antiglycolipid Antibodies ...... 387 5.3. Heterophil Antigens ...... 394 5.3.1. Forssman Antigen ...... 394 5.3.2. Hanganutziu-Deicher Antigen ...... 397 Contents xiii 5.4. Glycolipids with Blood Group ABH Specificities ...... 397 5.5. Glycolipids with Blood Group Lewis (Lea, Leb , Lec , and Led) Specificities ...... 404 5.5.1. Le and Leb Antigens ...... 405 5.5.2. LeC and Led Antigens...... 406 5.5.3. Le Antigens and X-Hapten Glycolipids ...... 407 5.6. Glycolipids with Blood Group P, Ph and pk Specificities ...... 407 5.7. Glycolipids with Blood Group I and i Specificities ...... 409 5.7.1. I and i Antigens and Antibodies ...... 409 5.7.2. Chemical Basis of I and i Specificities ...... 411 5.8. Glycolipids with Blood Group J-Antigen Specificity ...... 414 5.9. Tissue-Specific and Tumor-Associated Glycolipid Antigens ..... 415 5.9.1 Classical Studies ...... 415 5.9.2. Current Studies with Experimental Cancer and Human Cancer ...... 415 5.9.3. Lacto-N-neotetraosylceramide in NILpy Tumor ...... 416 5.9.4. Ganglio-N-triaosylceramide in Mice Sarcoma and Lymphoma ...... 416 5.9.5. Forssman Antigen in Human Cancer ...... 417 5.9.6. Glycolipid Antigen Specific for SV40-Transformed Hamster Tumors ...... 417 5.9.7. Human Melanoma Antigen Defined by Monoclonal Antibody ...... 419 5.9.8. Burkitt Lymphoma-Associated Antigen Defined by Monoclonal Antibody ...... 419 5.9.9. Monosialo Ganglioside of Human Colon Carcinoma Defined by a Specific Monoclonal Antibody ...... 420 5.9.10. Polyfucosylated Lactosaminolipids in Human Gastrointestinal, Lung, and Liver Adenocarcinoma ...... 420 5.10. Modification of Blood Group Antigens ...... 420 5.10.1. Deletion of A and B Determinants ...... 422 5.10.2. Lewis Fucolipids in Tumors ...... 423 5.10.3. Precursor Accumulation ...... 424 5.10.4. Blood Group Determinants in Human Tumors Foreign to the Host (Illegitimate Blood Group Antigens) ...... 424 5.11. Tissue-Specific or Organ-Specific Glycolipid Antigens ...... 426 5.11.1. Tissue-Specific or Organ-Specific Glycolipids ...... 426 5.11.2. Lymphoid-Cell-Type-Specific Glycolipids ...... 427 5.12. Glycolipid Antigens Associated with Autoimmune Processes .... 428 5.13. References ...... 429

Chapter 6 Glycosphingolipids as Receptors Julian N. Kanfer 6.1. Gangliosides and the Serotonin Receptor ...... 437 6.2. Gangliosides and Acetylcholine ...... 438 Contents 6.3. Gangliosides and Interferon ...... 439 6.4. Gangliosides and Bacterial Toxins ...... 440 6.4.1. Botulinum Toxin ...... 440 6.4.2. Tetanus Toxin...... 441 6.4.3. Cholera Toxin ...... 444 6.4.4. Other Toxins ...... 458 6.5. Gangliosides and Lymphocyte Markers ...... 459 6.5.1. Miscellaneous...... 462 6.6. Gangliosides and Glycoprotein Hormone Receptors ...... 462 6.6.1. Others ...... 465 6.7. Conclusions...... 465 6.8. References...... 467

Index ...... 473