Conn Neuropeptide Technology

Synthesis, Assay, Purification, and Processing
1. Auflage 2013
ISBN: 978-1-4832-5949-9
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, 440 Seiten

Methods in Neurosciences, Volume 6: Neuropeptide Technology: Synthesis, Assay, Purification, and Processing describes procedures and tools of assay useful for the identification, purification, and quantification of neuropeptides and their receptors. This volume is divided into four sections- chemical synthesis and biosynthesis; measurement of neuropeptides; purification and characterization; and neuropeptide degrading and processing enzymes. In these sections, this book specifically discusses the synthesis of peptide substrates for protein kinase C; synthesis of glycosyl neuropeptides; and ultrastructural localization of peptides. The measurement of neurokinin B by radioimmunoassay; purification and characterization of neuroendocrine peptides from rat brain; and preparation of glia maturation factor á are also elaborated. This text likewise covers the assays for arginine/lysine carboxypeptidases and enzymes that metabolize atrial natriuretic peptide. This publication is beneficial to neuroscientists and students researching on the synthesis, assay, purification, and processing of neuropeptides.

Autoren/Hrsg.

Conn, P. Michael

Inhaltsverzeichnis

1;Front Cover;1
2;Neuropeptide Technology: Synthesis, Assay, Purification, and Processing;4
3;Copyright Page;5
4;Table of Contents;6
5;Contributors to Volume 6;10
6;Preface;14
7;Methods in Neurosciences;16
8;Section I: Chemical Synthesis and Biosynthesis;18
8.1;Chapter 1. Large-Scale Synthesis of Gonadotropin-Releasing Hormone Antagonists for Clinical Investigations;20
8.1.1;Introduction;20
8.1.2;Nal-Glu Antagonist;21
8.1.3;Chemicophysical Characterization of Nal-Glu Antagonist and Azaline;36
8.1.4;Acknowledgments;42
8.1.5;References;42
8.2;Chapter 2. Synthesis of Peptide Substrates for Protein Kinase C;44
8.2.1;Introduction;44
8.2.2;Preparation of Peptide Substrate;45
8.2.3;Assay Method;46
8.2.4;Application to Crude Extract;48
8.2.5;Acknowledgments;51
8.2.6;References;51
8.3;Chapter 3. Synthesis of Glycosyl Neuropeptides;52
8.3.1;Introduction;52
8.3.2;Glycosylation Methodologies;52
8.3.3;General Comments;64
8.3.4;Reagents and Solvents;65
8.3.5;Acknowledgments;66
8.3.6;References;66
8.4;Chapter 4. Assays for Biosynthesis of Thyrotropin-Releasing Hormone;68
8.4.1;Introduction;68
8.4.2;Transfection of AtT20 Cell Line with PreproTRH cDNA;68
8.4.3;Pulse-Chase Studies of ProTRH-Derived Peptides;74
8.4.4;Appendix;81
8.4.5;Acknowledgment;85
8.4.6;References;85
8.5;Chapter 5. Microsequence Analysis of Proteins Purified by Gel Electrophoresis;86
8.5.1;Introduction;86
8.5.2;Outline of Procedure;87
8.5.3;Discussion;92
8.5.4;Experimental Procedures;93
8.5.5;Acknowledgments;100
8.5.6;References;101
8.6;Chapter 6. Preparation of Site-Specific Peptide Immunogens Using Multiple Antigen Peptide Approach System;102
8.6.1;Introduction;102
8.6.2;Multiple Antigen Peptide Design;103
8.6.3;Direct Approach;103
8.6.4;Indirect Approach;104
8.6.5;Experimental Procedures;107
8.6.6;Copy Number of Dendritic Arms;119
8.6.7;Relationship of Orientation and Antigenicity;119
8.6.8;Conclusion;122
8.6.9;Acknowledgments;123
8.6.10;References;123
9;Section II: Measurement of Neuropeptides;126
9.1;Chapter 7. Subcellular Distribution of Neuropeptides and Measurement of Their in Vitro Release;128
9.1.1;General Introduction;128
9.1.2;Subcellular Fractionation;128
9.1.3;Neuropeptide Release;131
9.1.4;References;135
9.2;Chapter 8. Analysis of Tachykinin Peptide Family Gene Expression Patterns by Combined High-Performance Liquid Chromatography-Radioimmunoassay;136
9.2.1;Preparation of Synthetic Tachykinin Peptides and Precursor Fragments;137
9.2.2;Extraction of Tachykinin Peptides from Tissues and Cells;142
9.2.3;Acknowledgments;146
9.2.4;References;146
9.3;Chapter 9. Ultrastructural Localization of Peptides: Comparison of Methods;147
9.3.1;Introduction;148
9.3.2;Methods;149
9.3.3;Results;154
9.3.4;Discussion;161
9.3.5;Acknowledgments;163
9.3.6;References;163
9.4;Chapter 10. Measurement of Immune System in Response to Peptides in Central Nervous System;164
9.4.1;Introduction;164
9.4.2;Methods;166
9.4.3;Results and Discussion;170
9.4.4;Acknowledgments;173
9.4.5;References;174
9.5;Chapter 11. Identification of Functionally Significant Phosphorylation Sites on Neuronal Proteins and Preparation of Antibodies That Recognize Them;175
9.5.1;Introduction;175
9.5.2;Identification of Functionally Significant Phosphorylation Sites;176
9.5.3;Preparation of Antibodies That Recognize Individual Phosphorylated Sites;184
9.5.4;Conclusions;191
9.5.5;Acknowledgments;192
9.5.6;References;192
9.6;Chapter 12. Measurement of Brain Peptides: Angiotensin and Atrial Natriuretic Peptide in Tissue and Cell Culture;194
9.6.1;Introduction;194
9.6.2;Methods;195
9.6.3;Cell Culture Methods;214
9.6.4;Acknowledgments;222
9.6.5;References;222
9.7;Chapter 13. Regionally Specific Antisera to Human ß-Preprotachykinin;224
9.7.1;Introduction;224
9.7.2;Radioimmunoassay for N-Terminal Flanking Peptide of Human /3-Preprotachykinin;226
9.7.3;Radioimmunoassay for Substance P Using Antiserum Directed against COOH-Terminal Residues;229
9.7.4;Radioimmunoassay for Neuropeptide K Using NH2-Terminally Directed Antiserum;231
9.7.5;Radioimmunoassay for Neuropeptide K(1-24) Peptide Using COOH-Terminally Directed Antiserum;232
9.7.6;Radioimmunoassay for Neurokinin A Using COOH-Terminally Directed Antiserum;234
9.7.7;Radioimmunoassay for COOH-Terminal Flanking Peptide of ß-Preprotachykinin;235
9.7.8;References;237
9.8;Chapter 14. Measurement of Neurokinin B by Radioimmunoassay;238
9.8.1;Introduction;238
9.8.2;Measurement of Neurokinin B by Radioimmunoassay Using NH2-Terminally Directed Antiserum;239
9.8.3;Measurement of Neurokinin B Radioimmunoassay Using COOH-Terminally Directed Antiserum;244
9.8.4;Comparison of Radioimmunoassay Methods for Measurement of Neurokinin B;247
9.8.5;References;248
9.9;Chapter 15. Radioimmunoassay of Tachykinins;249
9.9.1;Introduction;249
9.9.2;Preparation of Immunogens;249
9.9.3;Immunization;250
9.9.4;Labeling with Bolton-Hunter Reagent;251
9.9.5;Tissue Extraction;257
9.9.6;Radioimmunoassay;258
9.9.7;References;263
9.10;Chapter 16. Radioimmunoassay of a-Melanocyte-Stimulating Hormone;264
9.10.1;Introduction: Biochemistry and Physiology;264
9.10.2;Radioimmunoassay;266
9.10.3;References;273
9.11;Chapter 17. Radioimmunoassay of Atrial Natriuretic Peptide;277
9.11.1;Introduction;277
9.11.2;Materials and Methods;278
9.11.3;Results;280
9.11.4;Discussion;286
9.11.5;References
;287
9.12;Chapter 18. Radioimmunoassay of Cholecystokinin;288
9.12.1;Introduction;288
9.12.2;Radiolabeling and Tracer Purification;289
9.12.3;Radioimmunoassay Procedures;290
9.12.4;Measurement of Plasma Cholecystokinin;291
9.12.5;Measurement of Tissue Cholecystokinin;294
9.12.6;Summary;295
9.12.7;Acknowledgments;296
9.12.8;References;296
9.13;Chapter 19. Secretion and Radioimmunoassay of Somatostatin: In Vitro System;298
9.13.1;Introduction;298
9.13.2;General Aspects of Radioimmunoassay of Somatostatin;299
9.13.3;Somatostatin Secretion from Isolated Pancreatic Islets;302
9.13.4;Secretion of Somatostatin from Isolated Pancreatic Islets;305
9.13.5;References;309
10;Section III: Purification and Characterization;312
10.1;Chapter 20. Purification and Characterization of Immunoregulatory Peptides from Neuroendocrine Tissues: Suppressin as a Model;314
10.1.1;Introduction;314
10.1.2;Bioassay;314
10.1.3;Extraction and Purification of Suppressin;315
10.1.4;Verification of the Novelty of Suppressin;320
10.1.5;Summary;322
10.1.6;References;322
10.2;Chapter 21. Purification and Characterization of Neuroendocrine Peptides from Rat Brain: Prosomatostatin Isolation;323
10.2.1;Introduction;323
10.2.2;Methods Applied for Extraction and Purification of Prosomatostatin;330
10.2.3;Characterization of Peptides and Propeptides: General Considerations;333
10.2.4;Methods Applied in Characterization of Prosomatostatin;335
10.2.5;References;337
10.3;Chapter 22. Preparation of Glia Maturation Factor ß;338
10.3.1;Purification of Natural GMF-ß by Conventional Method;338
10.3.2;Purification of Natural GMF-ß by Immunoaffinity;343
10.3.3;Monitoring Purification Process;348
10.3.4;Activation and Bioassay of Pure GMF-ß;349
10.3.5;Characterization of Purified Products;350
10.3.6;References;354
10.4;Chapter 23. Detection and Purification of Thyrotropin-Releasing Hormone Precursor Peptides Using Antisera Generated against Synthetic Peptides;354
10.4.1;Introduction;354
10.4.2;Antibody Production;355
10.4.3;Radioimmunoassay;358
10.4.4;Assay;359
10.4.5;Peptide Purification;364
10.4.6;Acknowledgment;370
10.4.7;References;370
10.5;Chapter 24. Physiologic Roles of Peptides as Central Nervous System Transmitters;371
10.5.1;Introduction;371
10.5.2;Demonstrating Presence of Authentic Peptide within Relevant CNS Areas;373
10.5.3;Characterizing the Biological Actions of Peptides;375
10.5.4;Identifying CNS Site(s) of Action of Peptides;376
10.5.5;Evaluating Neuroanatomical Relationships between Peptide Immunoreactivity, Peptide Receptors, and Sites of Action;381
10.5.6;Monitoring Peptide Release from Neurons;381
10.5.7;Development and Application of Methods for Altering Synthesis, Release, and Receptor Actions of Peptides;383
10.5.8;Additional Considerations;384
10.5.9;Acknowledgments;385
10.5.10;References;385
11;Section IV: Neuropeptide Degrading and Processing Enzymes;388
11.1;Chapter 25. Assays for Arginine/Lysine Carboxypeptidases: Carboxypeptidases H (E; Enkephalin Convertase), M, and N;390
11.1.1;Introduction;390
11.1.2;Assay Methods for Arginine/Lysine Carboxypeptidases;390
11.1.3;Assay with Dns-Ala-Arg;396
11.1.4;Acknowledgment;401
11.1.5;References;401
11.2;Chapter 26. Assay of Peptidase That Removes Carboxy-Terminal Tripeptide from 125I-Labeled Atrial Natriuretic Factor;403
11.2.1;Introduction;403
11.2.2;Material and Methods;404
11.2.3;Discussion;412
11.2.4;References;413
11.3;Chapter 27. Assays for Enzymes That Metabolize Atrial Natriuretic Peptide;414
11.3.1;In Vitro Assay;415
11.3.2;Results and Discussion;416
11.3.3;Results and Discussion;421
11.3.4;References;425
11.4;Index;428

Leseproben

Contributors to Volume 6

Hiroaki Aihara,     (2), Department of Biochemistry, Kobe University School of Medicine, Kobe 650, Japan

Eduard Bardají,     (3), Chemistry Department, Loughborough University of Technology, Loughborough, Leicestershire LE11 3TU, England

Margery C. Beinfeld,     (7), Department of Pharmacological and Physiological Science, St. Louis University Medical Center, St. Louis, Missouri 63104

J. Edwin Blalock,     (20), Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, Alabama 35294

Marvin R. Brown,     (24), Departments of Medicine and Surgery, University of California, San Diego, San Diego, California 92103

J. Michael Conlon,     (13, 14), Department of Biomedical Sciences, Regulatory Peptide Center, Creighton University Medical School, Omaha, Nebraska 68178

J.D. Cremins,     (8), Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110

Lewis D. Fannon,     (10), Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 32610

Wolfgang H. Fischer,     (5), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Laurel A. Fisher,     (24), Department of Pharmacology, College of Medicine, University of Arizona Health Sciences Center, Tucson, Arizona 85724

Jon Florholmen,     (19), Department of Medicine and Institute of Clinical Medicine, University of Tromsø, Tromsø, Norway

Carolyn J. Foster,     (26), Department of Pharmacology, Schering-Plough Research, Bloomfield, New Jersey 07003

Nathan B. Fountain,     (9), Department of Anatomy, The University of Iowa, College of Medicine, Iowa City, Iowa 52242

Carl Hoeger,     (1), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Brent Jackson,     (5), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Ivor M.D. Jackson,     (4, 23), Division of Endocrinology, Brown University, Rhode Island Hospital, Providence, Rhode Island 02903

Evelyn Y. Jew,     (9), Department of Anatomy, The University of Iowa, College of Medicine, Iowa City, Iowa 52242

Jean Y. Jew,     (9), Department of Anatomy, The University of Iowa, College of Medicine, Iowa City, Iowa 52242

Gibbes R. Johnson,     (26), Division of Cytokine Biology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892

Janos Julesz,     (16), Endocrine Unit, First Department of Medicine, Albert Szent-Györgyi Medical University, H-6701 Szeged, Hungary

David Karr,     (5), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Mary B. Kennedy,     (11), Division of Biology, California Institute of Technology, Pasadena, California 91125

Birgitta Kimura,     (12), Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 32610

Dean Kirby,     (1), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Akira Kishimoto,     (2), Department of Biochemistry, Kobe University School of Medicine, Kobe 650, Japan

J.E. Krause,     (8), Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110

Philip A. Krieter,     (27), Department of Animal and Exploratory Drug Metabolism, Merck Sharp & Dohme Research Laboratories, Rahway, New Jersey 07065

Robert D. Leboeuf,     (20), Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, Alabama 35294

Ramon Lim,     (22), Division of Neurochemistry and Neurobiology, Department of Neurology, The University of Iowa, College of Medicine, Iowa City, Iowa 52242

Yi-An Lu,     (6), The Rockefeller University, New York, New York 10021

John E. Maggio,     (15), Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115

Charleen Miller,     (1), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Stephen G. Miller,     (11), Department of Cell and Developmental Biology, University of California, Berkeley, Berkeley, California 94720

Eduardo A. Nillni,     (4), Division of Endocrinology, Brown University, Rhode Island Hospital, Providence, Rhode Island 02903

Gillian M. Olins,     (27), Cardiovascular Diseases Research, Searle Research and Development, St. Louis, Missouri 63167

Minkyu Park,     (5), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Bruce L. Patton,     (11), Division of Biology, California Institute of Technology, Pasadena, California 91125

M. Ian Phillips,     (10, 12), Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 32610

Claudio Poiesi,     (17), Department of Chemistry, School of Medicine, University of Brescia, 25100 Brescia, Italy

John Porter,     (1), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Mohan K. Raizada,     (12), Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 32610

Roger D. Reidelberger,     (18), Department of Veterans Affairs Medical Center and Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, Nebraska 68105

Jean Rivier,     (1), The Clayton Foundation Laboratories for Peptide Biology, The Salk Institute, La Jolla, California 92037

Grace L. Rosenquist,     (18), Department of Animal Physiology, University of California, Davis, Davis, California 95616

Atsushi Sakurai,     (2), Department of Biochemistry, Kobe University School of Medicine, Kobe 650, Japan

Kevin A. Sevarino,     (4), Department of Psychiatry, Yale University School of Medicine, New Haven, Connecticut 06508

Randal A. Skidgel,     (25), Laboratory of Peptide Research, Departments of Pharmacology and Anesthesiology, College of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612

Günther Sperk,     (21), Department of Pharmacology, University of Innsbruck, A-6020...