Fain Lipid Metabolism in Signaling Systems

1. Auflage 2013
ISBN: 978-1-4832-8832-1
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, Band Volume 18, 357 Seiten

Reihe: Methods in Neurosciences

Many of the phospholipases and even the receptor for inositol 1,4,5-trisphosphate have recently been cloned and sequenced and play an important role in neurotransmission. Methods developed in recent years for the study of the enzymes and intermediates involved in the regulation of neuronal signal transduction involving phospholipid turnover are presented in this volume. - Techniques for examining the role of phosphoinositides in signal transduction - Techniques related to protein kinase C - Analysis of inositol phosphates - Techniques to analyze phospholipid turnover in the brain

Autoren/Hrsg.

Fain, John N.

Inhaltsverzeichnis

1;Front Cover;1
2;Lipid Metabolism in Signaling Systems;4
3;Copyright Page;5
4;Table of Contents;6
5;Contributors to Volume 18;10
6;Preface;16
7;Methods in Neurosciences;18
8;Section I: Phospholipases Involved in Signaling Systems;20
8.1;Chapter 1. Phosphatidylinositol Glycan-Anchor-Specific Phospholipase D from Mammalian Brain;22
8.1.1;Introduction;22
8.1.2;Assay Methods;23
8.1.3;Purification of Phosphoinositol Glycan-Specific Phospholipase D from Bovine and Human Brain;26
8.1.4;Characterization of Phosphoinositol Glycan-Specific Phospholipase D;28
8.1.5;References;31
8.2;Chapter 2. Rat Brain Membrane-Bound Phospholipase D;33
8.2.1;Introduction;33
8.2.2;Materials;34
8.2.3;Preparation of Rat Brain Membranes;34
8.2.4;Solubilization of Brain Membrane Phospholipase D Activity with Sodium Cholate;35
8.2.5;Assay of Solubilized Brain Membrane Phospholipase D Using C6-NBD-PC as Substrate;36
8.2.6;Properties of Cholate-Solubilized Phospholipase D Activity;37
8.2.7;Comments and Conclusions;42
8.2.8;Acknowledgments;43
8.2.9;References;43
8.3;Chapter 3. Soluble Phospholipase D;44
8.3.1;Introduction;44
8.3.2;Assay of Soluble Phospholipase D;44
8.3.3;Preparation of Cytosolic Enzyme;45
8.3.4;Properties of Cytosolic Enzyme;46
8.3.5;References;48
8.4;Chapter 4. Phosphatidylethanol Formation as Index of Phospholipase D Activity in Rat Brain Cortex Slices;49
8.4.1;Introduction;49
8.4.2;Experimental Procedures;51
8.4.3;Results and Comments;52
8.4.4;References;56
8.5;Chapter 5. Agonist Stimulation of Phosphoinositide Breakdown in Brain Membranes;57
8.5.1;Introduction;57
8.5.2;Assay with Membranes Made from [3H]inositol-Prelabeled Brain Slices;57
8.5.3;Assays with Exogenous [3H]Phosphoinositide Substrates;60
8.5.4;Assays with Membranes Directly Labeled with [3H]Inositol;63
8.5.5;Conclusion;66
8.5.6;References;66
8.6;Chapter 6. Regulation of Phosphoinositide Metabolism in Membranes Prepared from Human Brain Cortex;68
8.6.1;Introduction;68
8.6.2;Preparation of Membranes;69
8.6.3;Assay of Neurotransmitter Stimulation of Phosphoinositide-Specific Phospholipase C;70
8.6.4;Assays of Phosphatidylinositol Synthase and Phosphatidate Cytidylyltransferase;70
8.6.5;Assay of Phosphatidylinositol 4-Kinase;71
8.6.6;Results;72
8.6.7;Acknowledgments;75
8.6.8;References;75
8.7;Chapter 7. Purification of Phospholipase C-ß3 from Rat Brain;77
8.7.1;Introduction;77
8.7.2;Phospholipase C Assay;78
8.7.3;Phospholipase C-ß3 Purification Procedure;78
8.7.4;References;82
8.8;Chapter 8. Purification of Phospholipase C-ß4 and Phospholipase C-d2 from Bovine Retinas;83
8.8.1;Introduction;83
8.8.2;Materials;83
8.8.3;Phospholipase C Assay;83
8.8.4;Purification Procedure;83
8.8.5;References;90
8.9;Chapter 9. Purification of aq/11 from Brain;91
8.9.1;Introduction;91
8.9.2;Materials and Assays;91
8.9.3;Purification of aq/11;94
8.9.4;Reconstitution of Brain aq/11 with Phosphatidylinositol-Specific Phospholipase C;97
8.9.5;References;100
9;Section II: Other Enzymes Involved in Phospholipid Metabolism;102
9.1;Chapter 10. Purification of Phosphatidylinositol Synthase from Brain;104
9.1.1;Introduction;104
9.1.2;Preparation of Microsomes and Enzyme Solubilization;104
9.1.3;Assay Procedure for Solubilized Phosphatidylinositol Synthase;105
9.1.4;Purification of Solubilized Phosphatidylinositol Synthase on Cytidine Diphosphate Diacylglycerol Affinity Resin;105
9.1.5;Assay Procedure for Purified Phosphatidylinositol Synthase;108
9.1.6;Properties of Purified Phosphatidylinositol Synthase;109
9.1.7;Comments;110
9.1.8;Alternative Purification Approaches;110
9.1.9;References;111
9.2;Chapter 11. Phosphatidylinositol 4-Kinase from Bovine Brain;112
9.2.1;Introduction;112
9.2.2;Assay for Phosphatidylinositol 4-Kinase Activity;112
9.2.3;Enzyme Purification;113
9.2.4;Comments on Purification Procedures;117
9.2.5;References;118
9.3;Chapter 12. Phosphatidylinositol 3-Kinase;119
9.3.1;Introduction;119
9.3.2;Lipid Kinase Assays of Anti-P-Tyr Immunoprecipitated Proteins;121
9.3.3;Preparation of Protein A-Sepharose;125
9.3.4;Identification of D-3 Phosphorylated Lipids in Vivo in Ortho[32P]phosphate-Labeled Cells;125
9.3.5;Deacylation of Lipids;126
9.3.6;Chromatographic Separation of Deacylated Phospholipids (Glycerophosphoinositides);127
9.3.7;Identification of Phosphatidylinositol 3-Kinase p85 Protein Using Western Blotting Techniques and Enhanced Chemiluminescence;128
9.3.8;Acknowledgments;130
9.3.9;References;130
9.4;Chapter 13. Identification of Phosphatidylinositol Trisphosphate in Rat Brain;132
9.4.1;Introduction;132
9.4.2;Isolation of Inositol Phospholipids from Rat Brain;133
9.4.3;Analysis of Inositol Phospholipids;136
9.4.4;Concluding Remarks;141
9.4.5;Acknowledgments;142
9.4.6;References;143
10;Section III: Protein Kinase C;144
10.1;Chapter 14. Preparation of Protein Kinase C Isozymes and Substrates from Rat Brain;146
10.1.1;Introduction;146
10.1.2;Reagents;146
10.1.3;Assay of Protein Kinase C Activity;147
10.1.4;Preparation of Protein Kinase C Isozymes and Substrates from Rat Brain;149
10.1.5;Comments;153
10.1.6;References;155
10.2;Chapter 15. Protein Kinase C, a Zinc Metalloprotein: Quantitation of Zinc by Atomic Absorption Spectrometry;157
10.2.1;Introduction;157
10.2.2;Methods and Results of Zinc Measurements with Protein Kinase C;158
10.2.3;Concluding Remarks;170
10.2.4;Acknowledgment;171
10.2.5;References;172
10.3;Chapter 16. Expression of Protein Kinase C Isozymes in Insect Cells and Isolation of Recombinant Proteins;173
10.3.1;Introduction;173
10.3.2;Insect Cell Culture Techniques;174
10.3.3;Construction and Isolation of Recombinant Viruses;175
10.3.4;Detection of Recombinant Protein Kinase C Proteins, Solubility, and Phorbol Ester Binding;177
10.3.5;Expression and Purification of Classic Protein Kinase C Enzymes a, ßI, ßII, and .;179
10.3.6;Expression and Properties of Novel Protein Kinase C Enzymes;184
10.3.7;Posttranslational Modifications of Classic Group A Enzymes in Insect Cells;187
10.3.8;Comments;190
10.3.9;Acknowledgments;191
10.3.10;References;191
10.4;Chapter 17. Immunochemical Localization of Protein Kinase C and Phosphoinositide-Specific Phospholipase C;193
10.4.1;Introduction;193
10.4.2;Diacylglycerol Production and Protein Kinase Activation;194
10.4.3;Diacylglycerol Production and Protein Kinase Down-Modulation;197
10.4.4;Cytoskeletal Association of Protein Kinase C Isozymes;198
10.4.5;Protein Kinase C-Binding Proteins Target Protein Kinase C to Cytoskeleton;200
10.4.6;Nuclear Phosphoinositide-Specific Phospholipase C and Protein Kinase C;202
10.4.7;References;206
10.5;Chapter 18. Quantitative Analysis of Molecular Species of Diacylglycerol in Biological Samples;209
10.5.1;Introduction;209
10.5.2;Methods;210
10.5.3;Comments;216
10.5.4;References;217
11;Section IV: Measurement of Inositol Phosphates and Enzymes Regulating Inositol Phosphate Metabolism;218
11.1;Chapter 19. Measurement of Inositol Trisphosphate by Gas Chromatography/Mass Spectrometry: Femtomole Sensitivity Provided by Negative-Ion Chemical Ionization Mass Spectrometry in Submilligram Quantities of Tissue;220
11.1.1;Introduction;220
11.1.2;Procedures;221
11.1.3;Measurement of myo-Inositol and Validation of Analysis;226
11.1.4;Gas Chromatography/Mass Spectrometry Quantitation of Inositol in Tissue Samples;226
11.1.5;Summary;229
11.1.6;Acknowledgments;230
11.1.7;References;231
11.2;Chapter 20. Characterization of Inositol Phosphates by High-Performance Liquid Chromatography;232
11.2.1;Introduction;232
11.2.2;Principles;232
11.2.3;Experimental Procedure;233
11.2.4;Summary;241
11.2.5;Acknowledgments;241
11.2.6;References;241
11.3;Chapter 21. Measurement of Inositol 1,4,5-Trisphosphate, Inositol 1,3,4,5-Tetrakisphosphate, and Phosphatidylinositol 4,5-Bisphosphate in Brain;243
11.3.1;Introduction;243
11.3.2;Mass Assay of Inositol Phosphates;245
11.3.3;Radioreceptor Assays for Inositol 1,4,5-Trisphosphate;246
11.3.4;Use of Inositol 1,4,5-Trisphosphate Mass Assay for Phosphatidylinositol 4,5-Bisphosphate Determination;250
11.3.5;Radioreceptor Assay for Inositol 1,3,4,5-Tetrakisphosphate;251
11.3.6;Concluding Remarks;254
11.3.7;Appendix;255
11.3.8;Acknowledgments;260
11.3.9;References;261
12;Section V: Inositol Phosphate Receptors and Their Regulation;264
12.1;Chapter 22. Molecular Analysis of Inositol 1,4,5-Trisphosphate Receptors;266
12.1.1;Introduction;266
12.1.2;Purification of Inositol 1,4,5-Trisphosphate Receptors;267
12.1.3;Localization of Inositol 1,4,5-Trisphosphate Receptors;270
12.1.4;Cloning and Polymerase Chain Reaction Analysis of Inositol 1,4,5-Trisphosphate Receptor cDNAs;271
12.1.5;Structure–Function Relationships of Inositol 1,4,5-Trisphosphate Receptors;276
12.1.6;Molecular Basis of Inositol 1,4,5-Trisphosphate Receptor Regulation;280
12.1.7;How Do Inositol 1,4,5-Trisphosphate Receptors Work?;282
12.1.8;Acknowledgments;282
12.1.9;References;282
12.2;Chapter 23. Inositol 1,4,5-Trisphosphate Receptor Down-Regulation;285
12.2.1;Introduction;285
12.2.2;Quantification of Inositol 1,4,5-Trisphosphate-Induced Calcium Ion Mobilization in Control and Agonist-Pretreated Cells;285
12.2.3;Quantification of Inositol 1,4,5-Trisphosphate Receptor Concentration;293
12.2.4;Conclusions;297
12.2.5;Acknowledgment;297
12.2.6;References;297
12.3;Chapter 24. High-Affinity Inositol 1,3,4,5-Tetrakisphosphate Receptor from Cerebellum;299
12.3.1;Introduction;299
12.3.2;Buffers;300
12.3.3;Assay of Inositol 1,3,4,5-Tetrakisphosphate Binding Activity;300
12.3.4;Membrane Preparation;302
12.3.5;Receptor Solubilization;303
12.3.6;Purification of Inositol 1,3,4,5-Tetrakisphosphate Receptor;303
12.3.7;Identification of Inositol 1,3,4,5-Tetrakisphosphate Receptor;306
12.3.8;Properties of High-Affinity Inositol 1,3,4,5-Tetrakisphosphate Receptor;308
12.3.9;Photoaffinity Labeling of Inositol 1,3,4,5-Tetrakisphosphate Receptor;310
12.3.10;Modified Purification Scheme;313
12.3.11;Different Inositol 1,3,4,5-Tetrakisphosphate Receptors;314
12.3.12;Summary;315
12.3.13;Acknowledgments;315
12.3.14;References;315
12.4;Chapter 25. Inositol 1,4,5-Trisphosphate-Binding Proteins in Rat Brain Cytosol;317
12.4.1;Introduction;317
12.4.2;Binding Assay;318
12.4.3;Purification;318
12.4.4;Properties of Cytosolic Inositol 1,4,5-Trisphosphate-Binding Proteins;327
12.4.5;Conclusion;329
12.4.6;Acknowledgments;329
12.5;Chapter 26. Inositol 1,4,5-Trisphosphate Phosphatase and Kinase from Brain;331
12.5.1;Introduction;331
12.5.2;Assay of Enzymatic Activities;331
12.5.3;Preparation of Resins for Assay and Purification;333
12.5.4;Purification of Type I Inositol l,4,5-Trisphosphate-5-Phosphatase;334
12.5.5;Purification of Rat Brain Inositol l,4,5-Trisphosphate-3-Kinase;336
12.5.6;Acknowledgments;337
12.5.7;References;337
12.6;Chapter 27. Synthesis of 32P-Labeled Phosphoinositides and Inositol Phosphates: Characterization and Purification of Inositol 1,3,4,5-Tetrakisphosphate-3-phosphatase from Brain;339
12.6.1;Introduction;339
12.6.2;Preparation of Substrates;340
12.6.3;Assay for Inositol-1,3,4,5-Tetrakisphosphate-3-Phosphatase;348
12.6.4;Enrichment of Inositol-1,3,4,5-Tetrakisphosphate-3-Phosphatase from Porcine Brain;351
12.6.5;Properties of Inositol-1,3,4,5-Tetrakisphosphate-3-Phosphatase;352
12.6.6;Acknowledgments;354
12.6.7;References;354
12.7;Chapter 28. Species Differences in the Response of Second Messenger Inositol 1,4,5-Trisphosphate to Lithium;356
12.7.1;Introduction;356
12.7.2;Experimental Procedures;357
12.7.3;Results;359
12.7.4;Discussion;366
12.7.5;Summary;369
12.7.6;Acknowledgments;370
12.7.7;References;370
13;Index;372

Leseproben

Contributors to Volume 18
John C. Anthes(3),     Schering-Plough Research Institute, Kenilworth, New Jersey 07033 Elaine S.G. Bardes(15),     Section of Cell Growth, Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710 Robert M. Bell(15),     Section of Cell Growth, Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710 M. Motasim Billah(3),     Schering-Plough Research Institute, Kenilworth, New Jersey 07033 John Bloomenthal(15),     Section of Cell Growth, Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710 Roy A. Borchardt(15),     Section of Cell Growth, Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710 Urs Brodbeck(1),     Department of Neurochemistry, Institute of Biochemistry and Molecular Biology, University of Bern, CH-3012 Bern, Switzerland Lewis C. Cantley(12),     Division of Signal Transduction, Department of Medicine, Beth Israel Hospital, Boston, Massachusetts 02115 Vered Chalifa(2),     Department of Hormone Research, The Weizmann Institute of Science, Rehovot 76100, Israel R.A. John Challiss(21),     Department of Pharmacology and Therapeutics, University of Leicester, Leicester LE1 9HN, United Kingdom Enrique Claro(5),     Departamento de Bioquímica y Biología Molecular, Universidad Autónoma de Barcelona, E-08193 Bellaterra, Barcelona, Spain Michal Danin(2),     Department of Hormone Research, The Weizmann Institute of Science, Rehovot 76100, Israel John F. Dixon(28),     Department of Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53706 Joseph Eichberg(10),     Department of Biochemical and Biophysical Sciences, University of Houston, Houston, Texas 77204 Christophe Erneux(26),     Institut de Recherche Interdisciplinaire (IRIBHN), Université Libre de Bruxelles, B-1070 Brussels, Belgium John N. Fain(4, 5),     Department of Biochemistry, University of Tennessee, Center for Health Science, Memphis, Tennessee 38163 David Frith(16),     Max-Delbrück-Laboratorium, Max-Planck-Gesellschaft, D-50829 Köln, Germany Abdallah Ghalayini(10),     Department of Ophthamology, Baylor College of Medicine, Houston, Texas 77030 Amiya K. Hajra(18),     Mental Health Research Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48104 Masato Hirata(25),     Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka 812, Japan Marius C. Hoener(1),     Department of Neurochemistry, Institute of Biochemistry and Molecular Biology, University of Bern, CH-3012 Bern, Switzerland Ariane Höer(27),     Institut für Pharmakologie, Freie Universität Berlin, D-14195 Berlin, Germany Lowell E. Hokin(28),     Department of Pharmacology, University of Wisconsin Medical School, Madison, Wisconsin 53706 Fong-Fu Hsu(19),     Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110 Freesia L. Huang(14),     Section on Metabolic Regulation, Endocrinology and Reproduction Research Branch, NICHHD, National Institutes of Health, Bethesda, Maryland 20892 Kuo-Ping Huang(14),     Section on Metabolic Regulation, Endocrinology and Reproduction Research Branch, NICHHD, National Institutes of Health, Bethesda, Maryland 20892 Susan Jaken(17),     W. Alton Jones Cell Science Center, Lake Placid, New York 12946 Deok-Young Jhon(7),     Section on Signal Transduction, Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892 Takashi Kanematsu(25),     Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka 812, Japan David R. Kaplan(12),     Eukaryotic Signal Transduction Group, Molecular Mechanisms of Carcinogenesis Laboratory, ABL-Basic Research Program, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, Maryland 21702 Ryuichi Kato(20),     Department of Pharmacology, Keio University School of Medicine, Tokyo 160, Japan Karen Leach(17),     Department of Cell Biology, The Upjohn Company, Kalamazoo, Michigan 49001 Chang-Won Lee(8),     Section on Signal Transduction, Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892 Chunghee Lee(18),     Molecular Pathophysiology Branch, NIDDK, National Institutes of Health, Bethesda, Maryland 20892 Kweon-Haeng Lee(8),     Section on Signal Transduction, Laboratory of Biochemistry, NHLBI, National Institutes of Health, Bethesda, Maryland 20892 Mordechai Liscovitch(2),     Department of Hormone Research, The Weizmann Institute of Science, Rehovot 76100, Israel Marek Liyanage(16),     Max-Delbrück-Laboratorium, Max-Planck-Gesellschaft, D-50829 Köln, Germany Silvia Llahi(4),     Department of Biochemistry, University of Tennessee, Center for Health Science, Memphis, Tennessee 38163 Gregory A. Mignery(22),     Department of Physiology, Stritch School of Medicine, Loyola University Medical Center, Maywood, Illinois 60153 Heidi Möhn(2),     Department of Hormone Research, The Weizmann Institute of Science, Rehovot 76100, Israel Stefan R. Nahorski(21, 23),     Department of Pharmacology and Therapeutics, University of Leicester, Leicester LE1 9HN, United Kingdom Toshio Nakaki(20),     Department of Pharmacology, Keio University School of Medicine, Tokyo 160, Japan Eckard Oberdisse(27),     Institut für Pharmakologie, Freie Universität Berlin, D-14195 Berlin, Germany Ken-ichi Osada(14),     Section of Metabolic Regulation, Endocrinology and Reproduction Research Branch, NICHHD, National Institutes of Health, Bethesda, Maryland 20892 Dongeun Park(7),     Section on Signal Transduction, Laboratory of Biochemistry, NHLBI, National...