Methods of Biochemical Analysis: Analysis of Biogenic Amines and Their Related Enzymes, Supplement Volume

354 Pages·1971·15.19 MB·English

METHODS O F BIOCHEMICAL ANALYSIS
Supplement a1 Volume
W. E. COHN, Oak Ridge .Vational Laboratory, Tennessee
11. CONSUEN, The Canadian Red Cross Xemorial Hospital, l'aplow, Maidenhead,
Berkshire, England
J. GROSS, Department of Experimental Medicine and Cancer Research, Hebrew
University Hadassah Medical School, Jerusalern, Israel
H. HOLTEII, Carlsberg Laboratory, Copenhagen, Denmark
J. K. N. JONES, Deparlment of Organic Chemistry, Queen's University, Kingston,
Ontario, Canada
C. G. KING, Institute of AVutritiorL Sciences, Columbia University, New York,
New York
H. A. LARDY, Department of Biochemistry, University of Wisconsin, Madison
H. C. LICKSTEIN, Deparlment of .Ificrobiology, University of Cincinnati, Cin-
cinnati, Ohio
B. G. 3IALMSTRoM, Department of Biochemistry, University of Goteberg, Sweden
A. MEISTER, Department of Biochemistry, Cornell University Medical College,
New York, Sew York
0. NICKELSEN, Department of Food and .Vutrition, Michigan State University,
East Lansing, Michigan
J. ROCHE, Biochiniie Gbnhale el Comparbe, Collbge de France, Paris, Prance
A. TISELIUS, Institute of Biochemistry, University of Uppsala, Sweden
B. L. VALLEE, Biophysics Research Laboratory, Department of Biological Chemis-
try, Harvard Medical School, Boston, Jlassachusetls
I>. D. VAN SLYKE, Brookhaven .Vational Laboratory, Upton, Long Island, New
York
METHODS OF
BIOCHEMICAL ANALYSIS
ANALYSIS OF BIOGENIC AMINES AND
THEIR RELATED ENZYMES
Edited by DAVID GLICK
Stanford University Medical School
Stanf ord, Cal ijornia
SUPPLEMENTAL VOLUME
INTERSCIENCE PUBLISHERS a division of
John Wiley & Sons, Inc., New York London Sydney Toronto
The paper used in this book has a pH of 6.5 or higher. It has been used because the best
information now available indicates that this will contribute to its longevity.
Copyright @ 1971, by John Wilcy & Sons, Inc.
All Rights Reserved. Xo part of this book inay be rcproduccd by any means, nor
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of the publisher.
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Library of Congress Catalog Sumher 75-134036
IYBS 0-471-30420-4
PHISTED IS THE ITSITED Sl'.\'VES OF .\\IERIC.\
METHODS OF BIOCHEMICAL ANALYSIS SUPPLEMENTAL VOLUME
PREFACE
The interest in quantitative methods of measurement of biogenic amines
has followed from the remarkable advance in understanding the role of
these substances in many aspects of biology and medicine, but in some
cases this interest has led the advance by making possible means of inves-
tigation. However, whether leading or following, the influence of the
methodology, particularly on development of the neurosciences, has been
profound. In this highly active field advances in methods have been so
rapid that it is essential to provide critical evaluations from time to time
and to present selected procedures of special value.
The aim of this volume was to help meet the current need. The individ-
ual authors have had considerable direct experience with the methods they
treated, and they have provided practical analytical information on bio-
genic amines of particular interest and also on the enzymes involved in
their biosynthesis and degradation. In bringing this material together into
a single convenient source, I felt that the gain would be greater if the
authors were allowed more freedom to deal with their subjects as they
considered best, in spite of the Iack of uniformity and other advantages
that might accrue from a more rigidly set structure for the chapters.
Although this has indeed resulted in noticeable unevenness between indi-
vidual treatments, the minimization of restrictions has enabled the authors
to exploit their resources more effectively. In a few instances some minor
duplication of material among the chapters is evident, but it is minimal
and it was considered too unimportant to justify reorganization of the
writing for its elimination.
Ideas, concepts, and vision usually run far ahead of actually attainable
reality at a given time, so that the limiting factor in the advance of science
most often is the instrumentation and technique necessary to obtain
information-the methodology on which the experimental approach
depends. Thus it is hoped that this volume will contribute to the further
advance of the understanding of the biological and medical role of biogenic
amines.
DAVIDG LICK
V

METHODS OF BIOCHEMICAL ANALYSIS SUPPLEMENTAL VOLUME
CONTENTS
Assay of Serotonin, Related Metabolites, and Enzymes. By Walter
Lovenberg and Karl Engelman, Experimental Therapeutics
Branch, National Heart and Lung Institute, National Insti-
tutes of Health, Bethesda, Maryland. .................... 1
Determination of Amine Oxidases. By R. Kapeller-Adler, Depart-
ment of Pharmacology, University of Edinburgh Medical
School, Edinburgh, Scotland. ........................... 35
The Chemical Determination of Histamine. ByP . A.S hore, Depart-
ment of Pharmacology, University of Texas Southwestern
Medical School, Dallas, Texas. ......................... 89
Determination of Histidine Decarboxylase Activity. By Richard W.
Schayer, Research Center, Rockland State Hospital, Orange-
burg, New York... ................................... 99
The Chemical Estimation of Catecholamines and Their Metabolites
in Body Fluids and Tissue Extracts. By H. Weil-Malherbe,
Section on Neurochemistry, Division of Special Mental
Health Research, National Institute of Mental Health, St.
Elizabeths Hospital, Washington, D.C.. .................. 119
Assay of Enzymes of Catecholamine Biosynthesis and Metabolism.
By C. R. Creveling and J. W.D aly, Laboratory of Chemistry,
National Institute of Arthritis and Metabolic Diseases,
National Institutes of Health, Bethesda, Maryland. ........ 153
Measurement of Choline Esters. By Donald J. Jenden and L. B.
Campbell, UCLA School of Medicine and Brain Research
Institute, Los Angeles, California. ...................... 183
Determination of Activity of Cholinesterases. By Klas-Bertil
Augustinsson, Biochemical Institute, University of Stock-
holm, Sweden ........................................ 217
vii
...
Vlll CONTENTS
Measurement of Choline Acetylase. By J. Schuberth, Psychiatric
Research Center, Ulerdker Hospital, Uppsala, and B. Sorbo,
Division of Experimental Defence Afeclicine, Research Insti-
tute of National Defence, Sundbyberg, Sweden. ............ 275
Author Index ............................................... 297
Subject Index. .............................................. 319
Cumulative Index, Volumes 1-19 and Supplement. .............. 331
Methods of Biochemical Analysis, Supplemental Volume
Edited by David Glick
Copyright © 1971 by John Wiley & Sons, Inc.
Assay of Serotonin. Related Metabolites. and Enzymes
WALTER LOVENBERAGND KARLE NGELMAENx.p erimental Therapeutics Branch.
National Heart and Lung Institute. National Inslitutes of Health.
Bethesda. Maryland 20014
I . Introduction . . . . . . . . . . . . . . . . . . 1
I1. Methods of Detection and Quantitation . . . . . . . . . . 2
1. Spectrophotometric Analysis . . . . . . . . . . . . 4
A . Ultraviolet Absorption . . . . . . . . . . . . 4
B . Color Reactions of the 5-Hydroxyindole Nucleus . . . . . 4
2 . Fluorometric Analysis . . . . . . . . . . . . . . 6
3 . Chromatography . . . . . . . . . . . . . . . 9
I11. Isolation and Determination of 5-Hydroxyindoles . . . . . . . 12
. Serotonin . . . . . . . . . . . . . . . . . . 12
1
A . General Techniques for Tissue Assay . . . . . . . . 13
B . Blood Assay . . . . . . . . . . . . . . . 15
C . Urine . . . . . . . . . . . . . . . . . 18
.
2 5-Hydroxyindoleacetic Acid (5-HIAA) . . . . . . . . . 19
A . Urine Assay . . . . . . . . . . . . . . . 19
B . 5-HIAA in Tissues and Plasma . . . . . . . . . . 21
C . 5-HIAA in Cerebrospinal Fluid . . . . . . . . . . 22
3 . 5-Hydroxytryptophan . . . . . . . . . . . . . . 23
4 . Bufotenin (N,N' -Dimethylserotonin) . . . . . . . . . . 24
.
5 Melatonin (N-Acetyl-5-methoxytryptamine) . . . . . . . 25
fV. Enzymes Related to Serotonin Metabolism . . . . . . . . . 25
1. Tryptophan Hydroxylase . . . . . . . . . . . . . 25
A . Enzyme Preparation . . . . . . . . . . . . . 26
B . Reagents . . . . . . . . . . . . . . . . 26
C . Enzyme Assay Procedure . . . . . . . . . . . 27
2 . Aromatic >Amino Acid Decarboxylase . . . . . . . . . 28
A . Enzyme Preparation . . . . . . . . . . . . . 28
B . Enzyme . . . . . . . . . . . . . . . . . 30
.
3 Monoamine Oxidase . . . . . . . . . . . . . . . 30
4 . Hydroxyindole 0-Methyl Transferase . . . . . . . . . 31
A . Enzyme Preparation . . . . . . . . . . . . . 31
B . Reagents . . . . . . . . . . . . . . . . 32
C . Enzyme Assay Procedure . . . . . . . . . . . 32
References . . . . . . . . . . . . . . . . . . . . 32
.
I INTRODUCTION
It is now more than a decade since the previous review (81) was
published in this series. During this time severaI thousand scientific
articles have been published which have considered serotonin and have
reported measurements of either serotonin or its metabolities . Much of
I
2 WALTER LOVENBERG AND KARL ENGELMAN
this work utilized the methodology presented in the original review.
Several new or modified procedures have been developed and knowledge
of the enzymatic transformation of hydroxyindoles has increased. This
chapter attempts to integrate the new methods and point out situations
in which it is advantageous to use a particular procedure.
The huge literature which has appeared concerning serotonin indicates
that the biological scientist has been extremely fascinated with this com-
pound; yet, although it appears to be a neurohumoral agent, no unique or
specific physiological function can be assigned. Its pharmacological
effects have been studies in detail. Serotonin was first isolated from
beef serum in 1948 (57) as a vasoconstrictor substance, and identified as
5-hydroxytryptamine in 1949 (56). The material has since been found
widely distributed in nature. Many animal tissues contain significant
amounts of serotonin (25). It is notably present in brain and innervated
tissues, in blood platelets, and in intestinal mucosa. Many plants
also contain large amounts of 5-hydroxyindoles (79). Bananas contain
as much as 150 fig of serotonin per gram in peel and 30 pg/g in pulp.
Although the role of serotonin in plant physiology is unknown, its presence
in foods makes dietary restrictions essential in attempting to examine
man or animals for alterations in hydroxyindole metabolism.
The synthetic route for serotonin in plants is largely unexplored. In
animals however it is clear that serotonin is derived from the amino acid
tryptophan as shown in Fig. 1. The enzymes catalyzing the two reac-
tions, tryptophan hydroxylase and aromatic L-amino acid decarboxylase,
have been observed in numerous animal tissues. It is evident that
tryptophan hydroxylase is the rate-limiting factor in the conversion of
tryptophan to serotonin. The major serotonin metabolic pathway is
oxidation by monoamine oxidase and aldehyde dehydrogenase to yield
5-hydroxyindoleacetic acid (5-HIAA). In certain species however
appreciable amounts of conjugation occurs and large amounts of the
0-glucuronide or 0-sulfate are excreted in the urine after a loading dose of
serotonin. Another important pathway is the formation of melatonin in
the pineal gland. Melatonin content and the enzymes catalyzing its
formation undergo dramatic diurnal variations in pineal (4) and consider-
able attention has been devoted to its physiological role. We will attempt
to define the most suitable means for measuring the enzymes and the
reaction products in the 5-hydroxyindole pathway in the ensuing sections.
11. METHODS OF DETECTION AND QUANTITATION
Serotonin was first isolated on the basis of its biological activity,
and subsequently assayed in several bioassay systems (18,20,52,77).