Biochemical Actions of Hormones. Volume 9

364 Pages·1982·9.734 MB·English

by  Gerald Litwack (Eds.)

Biochemical Actions of Hormones
Edited by GERALD LITWACK
Fels Research Institute and Department of Biochemistry
Temple University, School of Medicine
Philadelphia, Pennsylvania
VOLUME IX
1982
ACADEMIC PRESS
A Subsidiary of Harcourt Brace Jovanovich, Publishers
New York London
Paris San Diego San Francisco Säo Paulo Sydney Tokyo Toronto
Contributors
DENNIS M. DISORBO TIIU OJASOO
HOWARD J. EISEN JACQUES POTTIER
ANDRÉ HERCHUELZ JEAN-PIERRE RAYNAUD
BRIAN B. HOFFMAN JEAN SALMON
ROBERT J. LEFKOWITZ S. STONEY SIMONS, JR.
FRANCES E. LELAND SANFORD S. SINGER
CHOH HAO LI DAVID A. SIRBASKU
GERALD LITWACK THOMAS C. SPELSBERG
WILLY J. MALAISSE ANDREW JOHN SZABO
THOMAS MICHEL OLGA SZABO
E. BRAD THOMPSON
COPYRIGHT © 1982, BY ACADEMIC PRESS, INC.
ALL RIGHTS RESERVED.
NO PART OF THIS PUBLICATION MAY BE REPRODUCED OR
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ACADEMIC PRESS, INC.
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United Kingdom Edition published by
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Library of Congress Cataloging in Publication Data
Main entry under title:
Biochemical actions of hormones.
Includes bibliographies and indexes.
1. Hormones—Collected works. I. Litwack, Gerald.
II. Axelrod, Julius, Date. [DNLM: 1. Hormones.
2. Physiology. WK102 B615]
QP571. B56 574.19·27 70-107567
ISBN 0-12-452809-0 (v. 9) AACR2
PRINTED IN THE UNITED STATES OF AMERICA
82 83 84 85 9 8 7 6 5 4 3 2 1
List of Contributors
Numbers in parentheses indicate the pages on which the authors' contributions begin.
Dennis M. DiSorbo (205), Fels Research Institute, and Department
of Biochemistry, Temple University, School of Medicine,
Philadelphia, Pennsylvania 19140
Howard J. Eisen (255), Developmental Pharmacology Branch, Na-
tional Institute of Child Health and Human Development, Na-
tional Institutes of Health, Bethesda, Maryland 20205
André Herchuelz (69), Laboratories of Experimental Medicine and
Pharmacology, Brussels University Medical School, Brussels,
Belgium
Brian B. Hoffman* (43), Departments of Medicine and Biochemistry,
The Howard Hughes Medical Institute, Duke University Medi-
cal Center, Durham, North Carolina 27710
Robert J. Lefkowitz (43), Departments of Medicine and Biochemis-
try, The Howard Hughes Medical Institute, Duke University
Medical Center, Durham, North Carolina 27710
Frances E. Leland (115), Departments of Biochemistry and Molecular
Biology, The University of Texas Medical School, Houston,
Texas 77025
Choh Hao Li (1), Hormone Research Laboratory, University of
California, San Francisco, California 94143
*Present address: Department of Medicine, Division of Clinical Pharmacology, Stanford
University Medical Center, Stanford, California 94305
IX
X List of Contributors
Gerald Litwack (205), Fels Research Institute and Department of
Biochemistry, Temple University, School of Medicine, Philadel-
phia, Pennsylvania 19140
Willy J. Malaisse (69), Laboratories of Experimental Medicine and
Pharmacology, Brussels University Medical School, Brussels,
Belgium B-1000
Thomas Michel (43), Departments of Medicine and Biochemistry,
The Howard Hughes Medical Institute, Duke University Medi-
cal Center, Durham, North Carolina 27710
Tiiu Ojasoo (305), Centre de Recherches Roussel-UCLAF, Paris
75007, France
Jacques Pottier (305), Centre de Recherches Roussel-UCLAF, Paris
75007, France
Jean-Pierre Raynaud (305), Centre de Recherches Roussel-UCLAF,
Paris 75007, France
Jean Salmon (305), Centre de Recherches Roussel-UCLAF, Paris
75007, France
S. Stoney Simons, Jr. (221), Laboratory of Chemistry, National Insti-
tute of Arthritis, Metabolism and Digestive Diseases, National
Institutes of Health, Bethesda, Maryland 20205
Sanford S. Singer (271), Chemistry Department, University of Day-
ton, Dayton, Ohio 45469
David A. Sirbasku (115), Departments of Biochemistry and Molecular
Biology, The University of Texas Medical School, Houston,
Texas 77025
Thomas C. Spelsberg (141), Department of Cell Biology, Section of
Biochemistry, Mayo Clinic, and Mayo Graduate School of
Medicine, Rochester, Minnesota 55901
Andrew John Szabo (93), Department of Medicine, New York Medi-
cal College, Valhalla, New York,and The Lincoln Hospital Medi-
cal Center, Bronx, New York 10451
Olga Szabo (93), Department of Medicine, New York Medical Col-
List of Contributors xi
lege, Valhalla, New York,and The Lincoln Hospital Medical
Center, Bronx, New York 10451
E. Brad Thompson (221), Laboratory of Biochemistry, National
Cancer Institute, National Institutes of Health, Bethesda, Mary-
land 20205
Preface
In this volume, the authors present a balanced overview of current re-
search on peptide and steroid hormones. C. H. Li opens the collection with
a review of the lipotropins and their active degradation products. An up-to-
date chapter on the regulation of adenylate cyclase by adrenergic receptors
follows by R. J. Lefkowitz and co-authors· T. Michel and B. B. Hoffman.
W. J. Malaisse and A. Herchuelz follow with an intriguing paper on potas-
sium ion conductance in the pancreatic beta cell. A. J. Szabo and O. Szabo
introduce a new and exciting concept of the insulin sensitive chemoreceptor of
the central nervous system. D. A. Sirbasku and F. E. Leland review their
new work on estrogen-induced growth factors with emphasis on their role in
tumor cell growth. T. C. Spelsberg reviews his excellent work on nuclear
acceptors for the avian progesterone receptor. J.-P. Raynaud presents a
cogent work on structural aspects of steroid hormones and their receptors,
whereas D. M. DiSorbo and G. Litwack review the current status of using
pyridoxal phosphate to provide a better understanding of the functions of
steroid receptors. S. S. Simons and E. B. Thompson review their elegant
work in producing a new affinity label for glucocorticoid receptors, and
H. Eisen reviews his ground-breaking work on the development of a poly-
clonal antibody to the glucocorticoid receptor. Finally, S. S. Singer appraises
us of his work on the regulation of the steroid sulfotransferases, a little-studied
but important new area.
The emphasis of this volume, then, has been on new developments along
several lines of investigation. I hope that future volumes will continue to
cover new developments as well as reviews of classical subjects undergoing
experimental revolutions.
Gerald Litwack
xiii
CHAPTER 1
The Lipotropins
Choh Hao Li
I. Introduction 1
II. Isohtion and Primary Structure of Sheep ß-LPH 2
///. Total Synthesis ofß -LPH 7
8
IV. A Glu1/ < Glu1 Polymorphism in ß-LPH 10
8
V. Isolation and Primary Structure of Human ß-LPH 12
A. Isolation of ß-LPH 14
h
B. Characterization of ß-LPH 15
h
C. Primary Structure of ß-LPH 17
h
VI. Isolation and Characterization ofy-LPH 18
VII. Lipotropins from Mouse Pituitary Cell 21
VIII. Conformation of ß-LPH 22
IX. Relationship of Structure to Lipolytic Activity of ß-LPH 23
8
X. Radioimmunoassay 24
XI. Immunohistochemical Localization of ß-LPH in the Pituitary and
Brain 25
XII. Biological Properties 27
XIII. ß-Lipotropin—the Prohormone for ß-Endorphin 29
XIV. y -Lipotropin—the Prohormone for ß-MSH 31
XV. Concluding Remarks 33
References 37
I. INTRODUCTION*
Pituitary extracts have been known for a long time to contain fat-
mobilizing or lipolytic activity (Best and Campbell, 1936). After pituitary
hormones have been isolated and their primary structures are known, it is
possible to conclude that somatotropin (GH), thyrotropin (TSH), corticotro-
* Abbreviations: LPH, lipotropin (subscript h denotes human; s, sheep; p, pig; b, bovine; and
t, turkey); CMC, carboxymethylcellulose; MSH, melanotropin; RIA, radioimmunoassay;
ACTH, corticotropin; EP, endorphin; CD, circular dichroism.
1
BIOCHEMICAL ACTIONS OF HORMONES, VOL. IX
Copyright © 1982 by Academic Press, Inc.
All rights of reproduction in any form reserved.
ISBN 0-12-452809-0
2 Choh Hao Li
pin (ACTH), and melanotropins (a- and /3-MSH) are active as lipolytic
agents. The discovery of a new pituitary lipolytic factor, whose chemical
characteristics are completely different from known pituitary hormones, oc-
curred in 1963 during the course of improving the procedure for the isolation
of ACTH from sheep pituitary glands. In 1955, we described a method for
the isolation of pure ACTH (Li et al., 1955) from acid-acetone extracts of
sheep pituitaries with a yield of only 9 mg from 1 kg of glands. In this
method, ACTH in the acid-acetone extract was adsorbed onto oxycellulose
before purification by means of countercurrent distribution and/or column
chromatography on ion-exchange resin. The oxycellulose adsorption step
does not remove ACTH completely from the extract and occasionally causes
inactivation of the hormone. For these reasons we omitted this step and
submitted the extract directly to chromatography on carboxymethylcel-
lulose. By this simplified procedure (Birk and Li, 1964), the yield of ACTH
increased considerably and, in addition, a new peak appeared before the
elution of ACTH. It was a matter of curiosity that the new peak was isolated
and purified. Physicochemical studies revealed it to possess distinct charac-
teristics that are different from those of ACTH and other known pituitary
hormones. During that time, we were actively investigating the structure-
activity relationship of synthetic peptides relating to the ACTH molecule
with regard to adrenocorticotropic, melanotropic, and lipolytic activities.
Among these three assay procedures, lipolytic assay was carried out more
frequently than the other two. Hence, the newly isolated component was
first submitted to lipolytic assay using the rabbit fat pads. It was found to be
active with a potency comparable to that of ACTH. Because of these assay
data, the name lipotropin (LPH) was proposed (Li, 1964) for the new prod-
uct. If melanotropic assay had been performed first, we might have called it
γ-melanotropin.
II. ISOLATION AND PRIMARY STRUCTURE OF SHEEP
0-LPH
The protocol for the isolation of sheep /3-LPH (/3-LPH) may be seen in
S
Table I (Li et al., 1966). The steps of acid-acetone extract, NaCl precipita-
tion, and dialysis were carried out at 0°C; the ACTH concentrate was
dialyzed against dilute NH OH (pH 8); and the insoluble material was cen-
4
trifuged off. The clear supernatant was lyophilized and yielded approxi-
mately 1 gm of fraction D' from 1 kg of whole or anterior pituitary glands.
Figure 1 presents a Chromatographie pattern of 1.5 gm. Fraction D' when
applied to a CMC column (1.5 x 60 cm) equilibrated with 0.01 M am-
monium acetate buffer of pH 4.6. Gradient elution (pH and buffer concen-
1. The Lipotropins 3
TABLE I
PROTOCOL FOR THE ISOLATION OF ß-LPH FROM SHEEP PITUITARY GLANDS
Yield
Procedure (gm)
Whole sheep pituitaries 1000
Acid-acetone extract" 25
NaCl precipitate0 4
Dialysis"; pH 8 soluble fraction* 1
CM-cellulose chromatography 0.08
IRC-50 chromatography 0.05
"These steps were carried out at 0°C.
6 Designated as fraction D'.
trations) was performed as previously described (Birk and Li, 1964). The
material corresponding to peak L, after lyophilization several times to re-
3
move ammonium acetate, was rechromatographed on a CMC column under
identical conditions as shown in Fig. 1. The yield of the lyophilized product
(L/) was approximately 80 mg from 1 kg of sheep pituitaries. Further purifi-
3
cation of L' was achieved on an IRC-50 (H+) column (2.2 X 26 cm) which
3
was previously equilibrated with 0.01 M ammonium acetate buffer of pH
4.6. One hundred milligrams of L/ in 10 ml starting buffer were applied to
3
the column and eluted with 1 M ammonium acetate buffer of pH 6.7; the
purified peptide, obtained from the main peak (II) as shown in Fig. 2, was
found to be homogeneous when again submitted to chromatography on
IRC-50 under the same conditions. The recovered product from the second
chromatography on IRC-50 was designated as /3-LPH. One hundred milli-
grams of/3-LPH were dissolved in 5 ml of 0.1 M NH HC0 and applied to a
4 3
Sephadex G-75 (Pharmacia, Uppsala) column (2.5 X 120 cm). The col-
umn was equilibrated with 0.1 M NH HC0 and had a hold-up volume of
4 3
100 ml. Elution was made with the same solvent. The peptide emerged as a
single peak with an elution volume of 158 ml. Thus, 50 mg pure )8-LPH
S
were obtained from 1 kg of whole sheep glands. It was also found that
acid-acetone extracts of posterior pituitaries contain no /3-LPH. This is in
agreement with the histochemical findings (Moon et al., 1973) that the
hormone is located in basophila of both the anterior and intermediate lobe
cells but not in the posterior lobe of the sheep pituitary.
/3 -LPH sediments in an ultracentrifuge as a single symmetrical peak at
S
59,780 rpm with so,w = 0.88 S. Sedimentation equilibrium studies at a
2
speed of 15,200 rpm with a peptide concentration of 0.5% gave an average
molecule weight for /3-LPH of 9500 using the calculated partial specified
S
volume of 0.728.