Angiogenesis In Vitro Systems In Vitro Systems

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Contributors xiii

Akiko Mammoto
Vascular Biology Program, Departments of Pathology & Surgery, Children’s
Hospital and Harvard Medical School, Boston, Massachusetts
Tadanori Mammoto
Vascular Biology Program, Departments of Pathology & Surgery, Children’s
Hospital and Harvard Medical School, Boston, Massachusetts
Manuela Martins-Green
Department of Cell Biology and Neuroscience, University of California, Riverside,
Riverside, California
Qing Robert Miao
Vascular Biology and Therapeutics Program and Department of Pharmacology,
Yale University School of Medicine, New Haven, Connecticut
William A. Muller
Department of Pathology, Northwestern University Feinberg School of Medicine,
Chicago, Illinois
Martin N. Nakatsu
Department of Molecular Biology and Biochemistry, University of California
Irvine, Irvine, California
Roberto F. Nicosia
Division of Pathology and Laboratory Medicine, Veterans Administration Puget
Sound Health Care System, Seattle, Washington, and Department of Pathology,
University of Washington, Seattle, Washington
Melissa Petreaca
Graduate Program in Cell, Molecular, and Developmental Biology, University of
California, Riverside, Riverside, California
Paramjeet K. Randhawa
Department of Biology, The University of North Carolina at Chapel Hill, Chapel
Hill, North Carolina
Cynthia A. Reinhart-King
Department of Biomedical Engineering, Cornell University, Ithaca, New York
Svetlana N. Rylova
Department of Biology, The University of North Carolina at Chapel Hill, Chapel
Hill, North Carolina
Anastasia Sacharidou
Department of Medical Pharmacology and Physiology, School of Medicine and
Dalton Cardiovascular Center, University of Missouri-Columbia, Columbia,
Missouri
xiv Contributors

Julia E. Sero
Vascular Biology Program, Departments of Pathology & Surgery, Children’s
Hospital and Harvard Medical School, Boston, Massachusetts
William C. Sessa
Vascular Biology and Therapeutics Program and Department of Pharmacology,
Yale University School of Medicine, New Haven, Connecticut
Masabumi Shibuya
Department of Molecular Oncology, Graduate School of Medicine and Dentistry,
Tokyo Medical and Dental University, Tokyo, Japan
Akio Shimizu
Vascular Biology Program, Department of Surgery, Children’s Hospital, Harvard
Medical School, Boston, Massachusetts
Peter T. C. So
Departments of Mechanical and Biological Engineering, Massachusetts Institute of
Technology, Cambridge, Massachusetts
Amber N. Stratman
Department of Medical Pharmacology and Physiology, School of Medicine and
Dalton Cardiovascular Center, University of Missouri-Columbia, Columbia,
Missouri
Erika S. Wittchen
Department of Cell and Developmental Biology, Lineberger Comprehensive
Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill,
North Carolina
Min Yao
Wellman Center for Photomedicine, Massachusetts General Hospital, Harvard
Medical School, Boston, Massachusetts
Penelope Zorzi
Department of Pathology, University of Washington, Seattle, Washington
PREFACE: A TRIBUTE TO DR. JUDAH FOLKMAN

The field of angiogenesis has recently lost its pioneer and leader, Dr. Judah
Folkman. This was a tremendous loss to many of us who knew him and the
field in general. Dr. Folkman inspired a generation of scientists lead an effort
to translate basic discovery toward new therapeutics for a wide range of
diseases including: cancer, blinding eye disease and inflammatory disease.
Due in large part to Dr. Folkman’s efforts and direction we now have the
first generation of therapeutics that disrupt angiogenesis in patients suffering
from cancer and macular degeneration. While Dr. Folkman clearly passed
away before his time, he did live long enough to observe that many
thousands of patients who are now better off due to anti-angiogenic therapy.
I had a rather interesting initiation to the field of anti-angiogenesis that
was completely inspired by Dr. Folkman. In the mid-1980s as a junior
faculty at the Scripps Research Institute I was studying what many of us in
the field were beginning to appreciate were a family of cell adhesion
receptors, later termed integrins. I had developed a monoclonal antibody
(LM609) to the vitronectin receptor later referred to as integrin anb3.
During the course of my work, LM609 was used to stain a variety of
diseased and normal tissues. To my surprise LM609 reacted strongly with
blood vessels in tumors and inflammatory sites but failed to react with blood
vessels in normal tissues. After seeing this result I began to read up on the
emerging field of angiogenesis research. It was clear that most of the
literature in the field came from Dr. Folkman or one of his disciples.
I immediately contacted Dr. Folkman. By the time I finished describing
our results I realized that he was excited as I was about our studies. In fact
before I could ask him any questions he suggested I come to his lab to learn
the chick chorioallantoic member (CAM) assay to determine whether
LM609 might have an impact on angiogenesis in a quantitative animal
model.
Naturally I arranged a trip to the Folkman lab within the next couple of
weeks. I had never been to Harvard and was a bit intimidated by the place.
As I took the elevator up to the Folkman lab and introduced myself to his
administrative assistant who welcomed me and indicated that Dr. Folkman
was expecting me. Within minutes Dr. Folkman, clad in a lab coat greated
me and suggested we get started. At this point I assumed he was going to
introduce me to one of his students or technicians who would then proceed
to show me the CAM assay step by step. To my surprise, Dr. Folkman lead
me to a hood, sat down and immediately started to instruct me how to

xv
xvi Preface

induce angiogenesis on the CAM. In fact, the next I knew I was sitting at
the hood next to Dr. Folkman going through the procedure in detail.
Therefore, I can say I learned the technique from the master. Ultimately,
Dr. Folkman introduced me to several members of the Folkman lab includ-
ing Drs. Donald Inber, Pat D’Amore and Mike Klagsburn. I remember how
enthusiastic and interactive each of these folks were. In fact, I am happy to
say that I still maintain close contact with these individuals and have had
many opportunities over the years to discuss science and reminisce about
the past. In fact, Don, Pat and have all kindly contributed chapters to
Methods in Enzymology volumes on Angiogenesis.
It was on my airplane ride home that I began to realize my career was
about to make a change in course. From that point forward I began to focus
on the role of adhesion receptors in angiogenesis and began to realize that
blocking angiogenesis with integrin antagonists could have a very impres-
sive impact on the growth of tumors in mice. Importantly, two of the agents
we developed, including humanized LM609 have shown clinical activity in
patients with late stage cancer.
Since my initiation to field I have since followed Dr. Folkman’s work
and have attended dozens of his lectures. Listening to a Folkman lecture is
like watching one of your favorite movies, you can watch it over and over
again and still find something interesting to think about. It was difficult for
anyone to attend his lecture and not come away excited about science in
general and angiogenesis in particular. The field of angiogenesis has matured
over the past 25 years due in large part to Dr. Folkman’s drive, enthusiasm,
perseverance, and kindness. Dr. Folkman’s leadership has helped to recruit
many scientists and physicians from the academic and private sectors to
focus on new approaches to develop inhibitors of angiogenesis.
In the early days there were a limited number of technological appro-
aches to measure and study angiogenesis. The CAM assay was among the
first quantitative approaches to measure the growth of newly forming blood
vessels. From this humble beginning the field has exploded and as a result
we now have a wide range of techniques approaches and animal models
designed to monitor and study the growth of new blood vessels in develop-
ment, tissue remodeling and disease. These methods are described in detail
in this volume by many of the current leaders of the field.

David A. Cheresh
METHODS IN ENZYMOLOGY

VOLUME I. Preparation and Assay of Enzymes

Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME II. Preparation and Assay of Enzymes
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME III. Preparation and Assay of Substrates
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME IV. Special Techniques for the Enzymologist
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME V. Preparation and Assay of Enzymes
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME VI. Preparation and Assay of Enzymes (Continued)
Preparation and Assay of Substrates
Special Techniques
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME VII. Cumulative Subject Index
Edited by SIDNEY P. COLOWICK AND NATHAN O. KAPLAN
VOLUME VIII. Complex Carbohydrates
Edited by ELIZABETH F. NEUFELD AND VICTOR GINSBURG
VOLUME IX. Carbohydrate Metabolism
Edited by WILLIS A. WOOD
VOLUME X. Oxidation and Phosphorylation
Edited by RONALD W. ESTABROOK AND MAYNARD E. PULLMAN
VOLUME XI. Enzyme Structure
Edited by C. H. W. HIRS
VOLUME XII. Nucleic Acids (Parts A and B)
Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE
VOLUME XIII. Citric Acid Cycle
Edited by J. M. LOWENSTEIN
VOLUME XIV. Lipids
Edited by J. M. LOWENSTEIN
VOLUME XV. Steroids and Terpenoids
Edited by RAYMOND B. CLAYTON

xvii
xviii Methods in Enzymology

VOLUME XVI. Fast Reactions

Edited by KENNETH KUSTIN
VOLUME XVII. Metabolism of Amino Acids and Amines (Parts A and B)
Edited by HERBERT TABOR AND CELIA WHITE TABOR
VOLUME XVIII. Vitamins and Coenzymes (Parts A, B, and C)
Edited by DONALD B. MCCORMICK AND LEMUEL D. WRIGHT
VOLUME XIX. Proteolytic Enzymes
Edited by GERTRUDE E. PERLMANN AND LASZLO LORAND
VOLUME XX. Nucleic Acids and Protein Synthesis (Part C)
Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN
VOLUME XXI. Nucleic Acids (Part D)
Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE
VOLUME XXII. Enzyme Purification and Related Techniques
Edited by WILLIAM B. JAKOBY
VOLUME XXIII. Photosynthesis (Part A)
Edited by ANTHONY SAN PIETRO
VOLUME XXIV. Photosynthesis and Nitrogen Fixation (Part B)
Edited by ANTHONY SAN PIETRO
VOLUME XXV. Enzyme Structure (Part B)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME XXVI. Enzyme Structure (Part C)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME XXVII. Enzyme Structure (Part D)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME XXVIII. Complex Carbohydrates (Part B)
Edited by VICTOR GINSBURG
VOLUME XXIX. Nucleic Acids and Protein Synthesis (Part E)
Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE
VOLUME XXX. Nucleic Acids and Protein Synthesis (Part F)
Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN
VOLUME XXXI. Biomembranes (Part A)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME XXXII. Biomembranes (Part B)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME XXXIII. Cumulative Subject Index Volumes I-XXX
Edited by MARTHA G. DENNIS AND EDWARD A. DENNIS
VOLUME XXXIV. Affinity Techniques (Enzyme Purification: Part B)
Edited by WILLIAM B. JAKOBY AND MEIR WILCHEK
Methods in Enzymology xix

VOLUME XXXV. Lipids (Part B)

Edited by JOHN M. LOWENSTEIN
VOLUME XXXVI. Hormone Action (Part A: Steroid Hormones)
Edited by BERT W. O’MALLEY AND JOEL G. HARDMAN
VOLUME XXXVII. Hormone Action (Part B: Peptide Hormones)
Edited by BERT W. O’MALLEY AND JOEL G. HARDMAN
VOLUME XXXVIII. Hormone Action (Part C: Cyclic Nucleotides)
Edited by JOEL G. HARDMAN AND BERT W. O’MALLEY
VOLUME XXXIX. Hormone Action (Part D: Isolated Cells, Tissues,
and Organ Systems)
Edited by JOEL G. HARDMAN AND BERT W. O’MALLEY
VOLUME XL. Hormone Action (Part E: Nuclear Structure and Function)
Edited by BERT W. O’MALLEY AND JOEL G. HARDMAN
VOLUME XLI. Carbohydrate Metabolism (Part B)
Edited by W. A. WOOD
VOLUME XLII. Carbohydrate Metabolism (Part C)
Edited by W. A. WOOD
VOLUME XLIII. Antibiotics
Edited by JOHN H. HASH
VOLUME XLIV. Immobilized Enzymes
Edited by KLAUS MOSBACH
VOLUME XLV. Proteolytic Enzymes (Part B)
Edited by LASZLO LORAND
VOLUME XLVI. Affinity Labeling
Edited by WILLIAM B. JAKOBY AND MEIR WILCHEK
VOLUME XLVII. Enzyme Structure (Part E)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME XLVIII. Enzyme Structure (Part F)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME XLIX. Enzyme Structure (Part G)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME L. Complex Carbohydrates (Part C)
Edited by VICTOR GINSBURG
VOLUME LI. Purine and Pyrimidine Nucleotide Metabolism
Edited by PATRICIA A. HOFFEE AND MARY ELLEN JONES
VOLUME LII. Biomembranes (Part C: Biological Oxidations)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
xx Methods in Enzymology

VOLUME LIII. Biomembranes (Part D: Biological Oxidations)

Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME LIV. Biomembranes (Part E: Biological Oxidations)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME LV. Biomembranes (Part F: Bioenergetics)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME LVI. Biomembranes (Part G: Bioenergetics)
Edited by SIDNEY FLEISCHER AND LESTER PACKER
VOLUME LVII. Bioluminescence and Chemiluminescence
Edited by MARLENE A. DELUCA
VOLUME LVIII. Cell Culture
Edited by WILLIAM B. JAKOBY AND IRA PASTAN
VOLUME LIX. Nucleic Acids and Protein Synthesis (Part G)
Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN
VOLUME LX. Nucleic Acids and Protein Synthesis (Part H)
Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN
VOLUME 61. Enzyme Structure (Part H)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME 62. Vitamins and Coenzymes (Part D)
Edited by DONALD B. MCCORMICK AND LEMUEL D. WRIGHT
VOLUME 63. Enzyme Kinetics and Mechanism (Part A: Initial Rate and
Inhibitor Methods)
Edited by DANIEL L. PURICH
VOLUME 64. Enzyme Kinetics and Mechanism
(Part B: Isotopic Probes and Complex Enzyme Systems)
Edited by DANIEL L. PURICH
VOLUME 65. Nucleic Acids (Part I)
Edited by LAWRENCE GROSSMAN AND KIVIE MOLDAVE
VOLUME 66. Vitamins and Coenzymes (Part E)
Edited by DONALD B. MCCORMICK AND LEMUEL D. WRIGHT
VOLUME 67. Vitamins and Coenzymes (Part F)
Edited by DONALD B. MCCORMICK AND LEMUEL D. WRIGHT
VOLUME 68. Recombinant DNA
Edited by RAY WU
VOLUME 69. Photosynthesis and Nitrogen Fixation (Part C)
Edited by ANTHONY SAN PIETRO
VOLUME 70. Immunochemical Techniques (Part A)
Edited by HELEN VAN VUNAKIS AND JOHN J. LANGONE
Methods in Enzymology xxi

VOLUME 71. Lipids (Part C)

Edited by JOHN M. LOWENSTEIN
VOLUME 72. Lipids (Part D)
Edited by JOHN M. LOWENSTEIN
VOLUME 73. Immunochemical Techniques (Part B)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 74. Immunochemical Techniques (Part C)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 75. Cumulative Subject Index Volumes XXXI, XXXII, XXXIV–LX
Edited by EDWARD A. DENNIS AND MARTHA G. DENNIS
VOLUME 76. Hemoglobins
Edited by ERALDO ANTONINI, LUIGI ROSSI-BERNARDI, AND EMILIA CHIANCONE
VOLUME 77. Detoxication and Drug Metabolism
Edited by WILLIAM B. JAKOBY
VOLUME 78. Interferons (Part A)
Edited by SIDNEY PESTKA
VOLUME 79. Interferons (Part B)
Edited by SIDNEY PESTKA
VOLUME 80. Proteolytic Enzymes (Part C)
Edited by LASZLO LORAND
VOLUME 81. Biomembranes (Part H: Visual Pigments and Purple Membranes, I)
Edited by LESTER PACKER
VOLUME 82. Structural and Contractile Proteins (Part A: Extracellular Matrix)
Edited by LEON W. CUNNINGHAM AND DIXIE W. FREDERIKSEN
VOLUME 83. Complex Carbohydrates (Part D)
Edited by VICTOR GINSBURG
VOLUME 84. Immunochemical Techniques (Part D: Selected Immunoassays)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 85. Structural and Contractile Proteins (Part B: The Contractile Apparatus
and the Cytoskeleton)
Edited by DIXIE W. FREDERIKSEN AND LEON W. CUNNINGHAM
VOLUME 86. Prostaglandins and Arachidonate Metabolites
Edited by WILLIAM E. M. LANDS AND WILLIAM L. SMITH
VOLUME 87. Enzyme Kinetics and Mechanism (Part C: Intermediates,
Stereo-chemistry, and Rate Studies)
Edited by DANIEL L. PURICH
VOLUME 88. Biomembranes (Part I: Visual Pigments and Purple Membranes, II)
Edited by LESTER PACKER
xxii Methods in Enzymology

VOLUME 89. Carbohydrate Metabolism (Part D)

Edited by WILLIS A. WOOD
VOLUME 90. Carbohydrate Metabolism (Part E)
Edited by WILLIS A. WOOD
VOLUME 91. Enzyme Structure (Part I)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME 92. Immunochemical Techniques (Part E: Monoclonal Antibodies and
General Immunoassay Methods)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 93. Immunochemical Techniques (Part F: Conventional Antibodies, Fc
Receptors, and Cytotoxicity)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 94. Polyamines
Edited by HERBERT TABOR AND CELIA WHITE TABOR
VOLUME 95. Cumulative Subject Index Volumes 61–74, 76–80
Edited by EDWARD A. DENNIS AND MARTHA G. DENNIS
VOLUME 96. Biomembranes [Part J: Membrane Biogenesis: Assembly and
Targeting (General Methods; Eukaryotes)]
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER
VOLUME 97. Biomembranes [Part K: Membrane Biogenesis: Assembly and
Targeting (Prokaryotes, Mitochondria, and Chloroplasts)]
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER
VOLUME 98. Biomembranes (Part L: Membrane Biogenesis: Processing
and Recycling)
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER
VOLUME 99. Hormone Action (Part F: Protein Kinases)
Edited by JACKIE D. CORBIN AND JOEL G. HARDMAN
VOLUME 100. Recombinant DNA (Part B)
Edited by RAY WU, LAWRENCE GROSSMAN, AND KIVIE MOLDAVE
VOLUME 101. Recombinant DNA (Part C)
Edited by RAY WU, LAWRENCE GROSSMAN, AND KIVIE MOLDAVE
VOLUME 102. Hormone Action (Part G: Calmodulin and
Calcium-Binding Proteins)
Edited by ANTHONY R. MEANS AND BERT W. O’MALLEY
VOLUME 103. Hormone Action (Part H: Neuroendocrine Peptides)
Edited by P. MICHAEL CONN
VOLUME 104. Enzyme Purification and Related Techniques (Part C)
Edited by WILLIAM B. JAKOBY
Methods in Enzymology xxiii

VOLUME 105. Oxygen Radicals in Biological Systems

Edited by LESTER PACKER
VOLUME 106. Posttranslational Modifications (Part A)
Edited by FINN WOLD AND KIVIE MOLDAVE
VOLUME 107. Posttranslational Modifications (Part B)
Edited by FINN WOLD AND KIVIE MOLDAVE
VOLUME 108. Immunochemical Techniques (Part G: Separation and
Characterization of Lymphoid Cells)
Edited by GIOVANNI DI SABATO, JOHN J. LANGONE, AND HELEN VAN VUNAKIS
VOLUME 109. Hormone Action (Part I: Peptide Hormones)
Edited by LUTZ BIRNBAUMER AND BERT W. O’MALLEY
VOLUME 110. Steroids and Isoprenoids (Part A)
Edited by JOHN H. LAW AND HANS C. RILLING
VOLUME 111. Steroids and Isoprenoids (Part B)
Edited by JOHN H. LAW AND HANS C. RILLING
VOLUME 112. Drug and Enzyme Targeting (Part A)
Edited by KENNETH J. WIDDER AND RALPH GREEN
VOLUME 113. Glutamate, Glutamine, Glutathione, and Related Compounds
Edited by ALTON MEISTER
VOLUME 114. Diffraction Methods for Biological Macromolecules (Part A)
Edited by HAROLD W. WYCKOFF, C. H. W. HIRS, AND SERGE N. TIMASHEFF
VOLUME 115. Diffraction Methods for Biological Macromolecules (Part B)
Edited by HAROLD W. WYCKOFF, C. H. W. HIRS, AND SERGE N. TIMASHEFF
VOLUME 116. Immunochemical Techniques
(Part H: Effectors and Mediators of Lymphoid Cell Functions)
Edited by GIOVANNI DI SABATO, JOHN J. LANGONE, AND HELEN VAN VUNAKIS
VOLUME 117. Enzyme Structure (Part J)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME 118. Plant Molecular Biology
Edited by ARTHUR WEISSBACH AND HERBERT WEISSBACH
VOLUME 119. Interferons (Part C)
Edited by SIDNEY PESTKA
VOLUME 120. Cumulative Subject Index Volumes 81–94, 96–101
VOLUME 121. Immunochemical Techniques (Part I: Hybridoma Technology and
Monoclonal Antibodies)
Edited by JOHN J. LANGONE AND HELEN VAN VUNAKIS
VOLUME 122. Vitamins and Coenzymes (Part G)
Edited by FRANK CHYTIL AND DONALD B. MCCORMICK
xxiv Methods in Enzymology

VOLUME 123. Vitamins and Coenzymes (Part H)

Edited by FRANK CHYTIL AND DONALD B. MCCORMICK
VOLUME 124. Hormone Action (Part J: Neuroendocrine Peptides)
Edited by P. MICHAEL CONN
VOLUME 125. Biomembranes (Part M: Transport in Bacteria, Mitochondria, and
Chloroplasts: General Approaches and Transport Systems)
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER
VOLUME 126. Biomembranes (Part N: Transport in Bacteria, Mitochondria, and
Chloroplasts: Protonmotive Force)
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER
VOLUME 127. Biomembranes (Part O: Protons and Water: Structure
and Translocation)
Edited by LESTER PACKER
VOLUME 128. Plasma Lipoproteins (Part A: Preparation, Structure, and
Molecular Biology)
Edited by JERE P. SEGREST AND JOHN J. ALBERS
VOLUME 129. Plasma Lipoproteins (Part B: Characterization, Cell Biology,
and Metabolism)
Edited by JOHN J. ALBERS AND JERE P. SEGREST
VOLUME 130. Enzyme Structure (Part K)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME 131. Enzyme Structure (Part L)
Edited by C. H. W. HIRS AND SERGE N. TIMASHEFF
VOLUME 132. Immunochemical Techniques (Part J: Phagocytosis and
Cell-Mediated Cytotoxicity)
Edited by GIOVANNI DI SABATO AND JOHANNES EVERSE
VOLUME 133. Bioluminescence and Chemiluminescence (Part B)
Edited by MARLENE DELUCA AND WILLIAM D. MCELROY
VOLUME 134. Structural and Contractile Proteins (Part C: The Contractile
Apparatus and the Cytoskeleton)
Edited by RICHARD B. VALLEE
VOLUME 135. Immobilized Enzymes and Cells (Part B)
Edited by KLAUS MOSBACH
VOLUME 136. Immobilized Enzymes and Cells (Part C)
Edited by KLAUS MOSBACH
VOLUME 137. Immobilized Enzymes and Cells (Part D)
Edited by KLAUS MOSBACH
VOLUME 138. Complex Carbohydrates (Part E)
Edited by VICTOR GINSBURG
Methods in Enzymology xxv

VOLUME 139. Cellular Regulators (Part A: Calcium- and

Calmodulin-Binding Proteins)
Edited by ANTHONY R. MEANS AND P. MICHAEL CONN
VOLUME 140. Cumulative Subject Index Volumes 102–119, 121–134
VOLUME 141. Cellular Regulators (Part B: Calcium and Lipids)
Edited by P. MICHAEL CONN AND ANTHONY R. MEANS
VOLUME 142. Metabolism of Aromatic Amino Acids and Amines
Edited by SEYMOUR KAUFMAN
VOLUME 143. Sulfur and Sulfur Amino Acids
Edited by WILLIAM B. JAKOBY AND OWEN GRIFFITH
VOLUME 144. Structural and Contractile Proteins (Part D: Extracellular Matrix)
Edited by LEON W. CUNNINGHAM
VOLUME 145. Structural and Contractile Proteins (Part E: Extracellular Matrix)
Edited by LEON W. CUNNINGHAM
VOLUME 146. Peptide Growth Factors (Part A)
Edited by DAVID BARNES AND DAVID A. SIRBASKU
VOLUME 147. Peptide Growth Factors (Part B)
Edited by DAVID BARNES AND DAVID A. SIRBASKU
VOLUME 148. Plant Cell Membranes
Edited by LESTER PACKER AND ROLAND DOUCE
VOLUME 149. Drug and Enzyme Targeting (Part B)
Edited by RALPH GREEN AND KENNETH J. WIDDER
VOLUME 150. Immunochemical Techniques (Part K: In Vitro Models of B and
T Cell Functions and Lymphoid Cell Receptors)
Edited by GIOVANNI DI SABATO
VOLUME 151. Molecular Genetics of Mammalian Cells
Edited by MICHAEL M. GOTTESMAN
VOLUME 152. Guide to Molecular Cloning Techniques
Edited by SHELBY L. BERGER AND ALAN R. KIMMEL
VOLUME 153. Recombinant DNA (Part D)
Edited by RAY WU AND LAWRENCE GROSSMAN
VOLUME 154. Recombinant DNA (Part E)
Edited by RAY WU AND LAWRENCE GROSSMAN
VOLUME 155. Recombinant DNA (Part F)
Edited by RAY WU
VOLUME 156. Biomembranes (Part P: ATP-Driven Pumps and Related Transport:
The Na, K-Pump)
Edited by SIDNEY FLEISCHER AND BECCA FLEISCHER