Table Of ContentHandbook of
Experimental Pharmacology
Continuation of Handbuch der experimentellen Pharmakologie
Vol. 66/1
Editorial Board
G. V. R. Born, London· A. Farah, Rensselaer, New York
H. Herken, Berlin· A. D. Welch, Memphis, Tennessee
Advisory Board
S. Ebashi· E. G. Erdos' V. Erspamer' U. S. von Euler' W S. Feldberg
G. B. Koelle' M. Rocha e Silva' 1. R. Vane' P. G. Waser
Glucagon I
Contributors
T. T. Aoki . T. L. Blundell . C. Bordi . 1. Brange . W W Bromer
G. F. Cahill, lr.· A. D. Cherrington' 1.-L. Chiasson· T.H. Claus
1. W Ensinck . S. Falkmer . D. W Foster· L. G. Heding
C. Hellerstrom . B. de Hemptinne . 1. 1. Holst . K. S. Koch
P.l. Lad . P.l. Lefebvre' H. L. Leffert· 1. D. McGarry
R.B. Merrifield· A.1. Moody· S. Mojsov . R. Nosadini
L. Orci . C. R. Park . A. Perrelet . S.1. Pilkis . M. Pingel
M. Rodbell . E. Samols . B. Skelbaek-Pedersen . H. Skelly
R.1. Smith· W. Stalmans . L. Thim . A. Tiengo . R. H. Unger
I. Valverde' S. Van Noorden
Editor
P.J. Lefebvre
Springer-Verlag
Berlin Heidelberg New York Tokyo 1983
Professor PIERRE 1. LEFEBVRE, M.D.
Professor of Medicine, University of Liege
Guest Professor, University of Brussels (V.U.B.)
Corresponding Member, Royal Academy of Medicine of Belgium
Chairman, Medical Policlinics, University of Liege
Head, Division of Diabetes, University of Liege
Institut de M6decine, Hopital de Baviere
Boulevard de la Constitution, 66
4020 Liege, Belgium
With 136 Figures
ISBN -13: 978-3-642-68868-3 e-ISBN-13: 978-3-642-68866-9
DOl: 10.1007/978-3-642-68866-9
Library of Congress Cataloging in Publication Data. Main entry under title: Glucagon. (Handbook of experimental
pharmacology; v. 66) Bibliography: p. Includes index. I. Glucagon -Addresses, essays, lectures. I. Lefebvre, Pierre J.
II. Series. [DNLM: I. Glucagon. WI HA51L vol. 66 pt. 1-2/WK 801 G5656] QP905.H3 vol. 66 [QP572.G5],
615'.ls [612'.34] 83-583
This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned,
specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying
machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are
made for other than private use, a fee is payable to "Verwertungsgesellschaft Wort", Munich.
© by Springer-Verlag Berlin Heidelberg 1983.
Softcover reprint of the hardcover 1st edition 1983
The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific
statement, that such names are exempt from the relevant protective laws and regulations and therefore free for
general use.
Product Liability: The publisher can give no guarantee for information about drug dosage and application thereof
contained in this book. In every individual case the respective user must check its accuracy by consulting other
pharmaceutical literature.
2122/3130-543210
List of Contributors
T. T. AOKI, Joslin Diabetes Center, One Joslin Place, Boston, MA 02215, USA
T. L. BLUNDELL, Laboratory of Molecular Biology, Department of Crystallo
graphy, Birkbeck College, University of London, Malet Street, London
WCIE 7HX, Great Britain
C. BORDI, Istituto di Anatomia ed Istologia Patologica, Universita di Parma,
Parma, Italy
J. BRANGE, Novo Research Institute, Novo Allee, 2880 Bagsvaerd, Denmark
W. W. BROMER, Research Advisor, Lilly Research Laboratories, Indianapolis,
IN 46285, USA
G. F. CAHILL, Jr., Director of Research, Howard Hughes Medical Institute,
398 Brookline Avenue, Suite 8, Boston, MA 02215, USA
A. D. CHERRINGTON, Department of Physiology, Howard Hughes Medical
Institute, Vanderbilt University, 702 Light Hall, Nashville, TN 37232, USA
J.-L. CHIASSON, Director, Research Laboratory in Diabetes and Carbohydrates
Metabolism, Clinical Research Institute of Montreal, 110 Avenue des Pins
Ouest, Montreal, Que. H2W lR7, Canada
T. H. CLAUS, American Cyanamid Company, Medical Research Division, Lederle
Laboratories, Pearl River, NY 10965, USA
J. W. ENSINCK, Department of Medicine, Program Director, Clinical Research
Center, University Hospital, RC-14, University of Washington, Seattle,
WA 98195, USA
S. FALKMER, Department of Pathology, Malmo General Hospital, 21401 Malmo,
Sweden
D. W. FOSTER, Department of Internal Medicine, University of Texas Health
Science Center at Dallas, Southwestern Medical School, 5323 Harry Hines
Boulevard, Dallas, TX 75235, USA
L. G. HEDING, Medical Department, Novo Research Institute, Novo Allee,
2880 Bagsvaerd, Denmark
C. HELLERSTROM, University of Uppsala, Department of Medical Cell Biology,
Biomedicum, Box 571, 75123 Uppsala, Sweden
VI List of Contributors
B. DE HEMPTINNE, Laboratory of experimental Surgery, Catholic University of
Louvain, 1200 Brussels, Belgium
J. J. HOLST, Institute of Medical Physiology C, University of Copenhagen, The
Panum Institute, Blegdamsvej 3,2200 Copenhagen N, Denmark
K. S. KOCH, Department of Medicine, M - 013 H, Division of Pharmacology,
School of Medicine, University of California at San Diego, La Jolla, CA 92093,
USA
P. J. LAD, Department of Medicine, M - 013 H, Division of Pharmacology,
School of Medicine, University of California at San Diego, La Jolla, CA 92093,
USA
P. J. LEFEBVRE, Head of the Division of Diabetes, Universite de Liege, Hopital
Universitaire de Baviere, Institut de Medecine, Diabetologie, Boulevard de la
Constitution, 66, 4020 Liege, Belgium
H. L. LEFFERT, Department of Medicine, M-Ol3 H, Division of Pharmacology,
School of Medicine, University of California at San Diego, La Jolla, CA 92093,
USA
J. D. MCGARRY, Departments ofInternal Medicine and Biochemistry, University
of Texas Health Science Center at Dallas, Southwestern Medical School,
5323 Harry Hines Boulevard, Dallas, TX 75235, USA
R. B. MERRIFIELD, The Rockefeller University, 1230 York Avenue, New York,
NY 10021, USA
A. J. MOODY, Novo Research Institute, Novo Allee, 2880 Bagsvaerd, Denmark
S. MOJsov, The Rockefeller University, 1230 York Avenue, New York, NY 10021,
USA
R. NOSADINI, Istituto di Medicina Clinica dell'Universita di Pad ova, Via
Giustiniani, 2, 35100 Padova, Italy
L. ORCI, Institute of Histology and Embryology, University of Geneva Medical
School, 1211 Geneva 4, Switzerland
C. R. PARK, Chairman, Department of Physiology, Vanderbilt University, School
of Medicine, Nashville, TN 37232, USA
A. PERRELET, Institute of Histology and Embryology, University of Geneva,
Medical School, 1211 Geneva 4, Switzerland
S. J. PILKIS, Department of Physiology, Vanderbilt University, School of Medicine,
Nashville, TN 37232, USA
M. PINGEL, Novo Research Institute, Novo Allee, 2880 Bagsvaerd, Denmark
M. RODBELL, Section on Membrane Regulation, National Institute of Arthritis,
Diabetes, Digestive, and Kidney Diseases, Bethesda, MD 20205, USA
List of Contributors VII
E. SAMOLS, Division of Endocrinology, Metabolism and Radionuclide Studies,
Department of Medicine, V A Medical Center, 800 Zorn Avenue, Louisville,
KY 40202, USA
B. SKELBAEK-PEDERSEN, Novo Research Institute, Novo Allee, 2880 Bagsvaerd,
Denmark
H. SKELLY, Department of Medicine, M - 013 H, Division of Pharmacology,
School of Medicine, University of California at San Diego, La Jolla, CA 92093,
USA
R. J. SMITH, Howard Hughes Medical Institute, Laboratories at the Joslin Diabetes
Center, One Joslin Place, Boston, MA 02215, USA
W. STALMANS, Afdeling Biochemie, Faculteit Geneeskunde, Katholieke
Universiteit Leuven, Herestraat 49, 3000 Leuven, Belgium
L. TRIM, Novo Research Institute, Novo Allee, 2880 Bagsvaerd, Denmark
A. TIENGO, Istituto di Medicina Clinica dell'Universita di Padova, Cattedra di
Malattie del Ricambio, Via Giustiniani, 2, 35100 Padova, Italy
R. H. UNGER, Senior Medical Investigator, Dallas VA Medical Center, 5323 Harry
Hines Boulevard, Dallas, TX 75235, USA
I. VALVERDE, Fundaci6n Jimenez Diaz, U niversidad Aut6noma de Madrid, A vda.
Reyes Cat6licos 2, Ciudad Universitaria, Madrid 3, Spain
S. VAN NOORDEN, Department of Histopathology, Royal Postgraduate Medical
School, Hammersmith Hospital, London, Great Britain
Preface
The Editorial Board of the Handbook of Experimental Pharmacology apparently
did not hurry in suggesting production of a volume on glucagon since the present
opus is number sixty-six in the series. This fact is even more striking if we consider
that 34 volumes published over about eight years will separate the books on
glucagon from those on insulin on library shelves, whereas only a few microns
separate the cells manufacturing these two polypeptides within the islets of
Langerhans in the pancreas!
Numerous factors have probably caused this dicrimination; four of them are:
First, insulin deficiency or resistance is the cause of one of the most serious and
distressing diseases, diabetes mellitus, which affects millions of people, whereas
glucagon deficiency is apparently an extremely rare disorder, for which detailed
reports are published of individual cases whenever they occur.
Second, since its discovery in 1921 by BANTING and BEST, insulin has been
irreplaceable for the treatment of the most severe forms of diabetes, whereas, in
contrast, glucagon was until recently considered a relatively minor therapeutic
agent.
Third, whereas insulin is a compound which has been well characterized since the
pioneering work of SANGER and its biosynthesis clearly identified by STEINER and
his co-workers, glucagon, also well characterized chemically, has suffered from its
parenthood with the so-called "glucagon-like immunoreactive substances", an
incompletely defined series of immunologically related polypeptides present in the
gut, the pancreas and some other parts of the body.
Fourth, while the insulin-producing cell, the B-cell of the islets of Langerhans, has
received a great amount of attention from morphologists for many years, the
glucagon-producing cell or A-cell has long been considered as the "parent pauvre."
The credentials of glucagon are numerous however. It was among the very first
polypeptide hormones to be isolated, purified, sequenced, and synthetized. Thanks
to the efforts of UNGER, it was the second polypeptidic hormone to become
measurable by radioimmunoassay, a few months after insulin. It has served as a
very valuable tool which permitted SUTHERLAND and his co-workers to discover
cyclic AMP and RODBELL and his associates to investigate cell membrane receptors.
Our knowledge of physiology of glucagon is based on the work of STAHL, FOA,
SUTHERLAND, and DE DuVE, to mention only few of the pioneers, and on work
performed since the late fifties in Dallas by UNGER and his co-workers. In the
seventies UNGER and ORCI, combining their physiological and morphological
x Preface
expertises, renewed the study of the pathopysiology of diabetes; they considered the
microanatomy of the islets of Langerhans and demonstrated both the interaction
and the partition of the various cell types involved.
When asked to serve as editor for a volume on glucagon in the present series, I
immediately decided to accept since there was a great need for a comprehensive
book on this subject. As a matter of fact, my task of selecting and contacting
contributors has been easy: all are personal friends, all are undisputed experts in
their field, and all accepted my invitation.
The only major problem arose when all the contributions had been collected,
namely the total amount of pages exceeded by far that which could be reasonably
gathered in a single volume. Analysis of the content showed that shortening the
individual contributions was definitely unacceptable. Another possibility was to
renounce the comprehensive character of the book by leaving out several topics,
a position which was acceptable neither to the authors, who had already written
the chapters, nor to the editor, who had indicated to the contributors that
comprehensiveness was a major characteristic of the whole project. The help came
from the publisher, who accepted a third possible solution, that of producing
Glucagon in two volumes.
The first volume gathers the more "basic" contributions on the chemistry and
physicochemistry of glucagon, the morphology of the A-cells, the biosynthesis of
the hormone, its production, its immunogenicity, and its assay procedures. It also
contains ten chapers describing the actions of the hormone at the molecular level.
The second volume deals with the various factors controlling glucagon secretion,
the question of extrapancreatic glucagon, the place of glucagon in physiology and
in pathology, the catabolism of the hormone, the pharmacological effects of
glucagon, and its use in diagnosis and in therapy.
Two critiques often made on a work like this are redundancy and not being up
to date. We have done our best to eliminate both.
About redundancy, great efforts have been made to delineate the topics to be
treated by the various contributors, and direct contacts have been encouraged
between contributors having to deal with closely related topics. At the editorial
level, redundant paragraphs or figures have been deleted. Yet, it has been our
deliberate policy to preserve homogeneity of each chapter and, as a consequence,
some topics have been considered in two chapters on a different scope. For instance,
glicentin, the newly recognized precursor of glucagon, has been considered both in
the chapter on the biosynthesis of glucagon and in the one dealing with the various
glucagon-related peptides. Similarly, the chapter on "glucagon and liver output
in vivo" is a physiological review of mechanisms dealt with in a more biochemical
manner in the contributions on "liver glycogen metabolism" and on "gluconeo
genesis." With such an approach, we think that the reader will find both detailed
analyses of the mechanisms involved and comprehensive and integregated views on
the phenomena as they occur in the organism as a whole.
About up-dating, producing a multiauthored book like this takes time; this is
not, however, an excuse for it to be out of date when in print. Every effort has been
made by the contributors and by the editor to cover the literature completely up to
the very last possible moment. Newly published data have been introduced, when
Preface XI
necessary, at the various steps of scientific editing, copy editing, and galley-proof
correcting. We hope that this effort will be appreciated by the readers.
I wish to thank all those who participated in this enterprise: the authors for their
comprehensive contributions and their willingness to abide by the rules of the game,
my secretary Mrs. V AESSEN-PETIT in Liege who helped me in the editorial process,
Mr. EMERSON, in Great Britain, whose copy editing was outstanding and the
publisher, in Heidelberg, from whom I have received help from Mr. BERGSTEDT,
Mrs. WALKER, and Mr. BISCHOFF.
Our readers will be the judge of our joint efforts; let us hope that they will not
be disappointed.
PIERRE J. LEFEBVRE
Contents
Chemistry and Physicochemistry of Glucagon
CHAPTER 1
Chemical Characteristics of Glucagon. W. W. BROMER. With 2 Figures
A. Introduction. . . . . . 1
B. Isolation and Purification 1
C. Properties. . . . . . . 2
D. Amino Acid Sequence 3
E. Covalent Chemical Modification and Biologic Function 5
I. Limitations of the Approach . . . . 5
II. General Considerations . . . . . . . . . . 7
III. The Question of Several Active Sites. . . . . 7
IV. Amino Terminal and Diamino Modifications. 12
V. Inhibitors of Glucagon . . . . . . . . . . 13
VI. Modifications of Glutamyl, Lysyl, Arginyl, and Tryptophyl
Residues. . . . . . . . . . . . . . . . . . . . . . 14
VII. Modifications of Tyrosyl Residues . . . . . . . . . . 15
VIII. Methionyl Residue and Carboxyl Terminal Modifications 16
IX. Summary of Covalent Modifications and Function 18
References . . . . . . . . . . . . . . . . . . . . . . . . . 19
CHAPTER 2
The Chemical Synthesis of Glucagon. R. B. MERRIFIELD and S. MOJsov
With 8 Figures
A. Introduction. . . . . . 23
B. Early Synthetic Efforts . 24
C. The First Total Synthesis 24
D. Further Syntheses by Fragment Condensation in Solution 26
E. Solid Phase Fragment Synthesis 28
F. Stepwise Solid Phase Synthesis. 30
G. Conclusions 33
References . . . . . . . . . . . 34
