Methods of Biochemical Analysis, Volume 26

455 Pages·1980·13.724 MB·English

METHODS OF BIOCHEMICAL ANALYSIS
26
Volume
Advisory Board
N. G. ANDERSON, Divisum OJ BioIogual and Medical Research, Argonne National
hboratones, Illanok
TH. BUC H ER. lnstitute ofPhysiologua1 Chemictry, and Physical Bwchemistry and Cell Biology,
University of Munich, West C-ny
W. E. COHN, Oak Rdge National hboralory, Tennessee
P. DOUZOU, Institute o f Physizo-Chemical Biology, Edmond de Rothcchild Foundation, Paris,
France
R. W.E STABROOK, Department oJBiochemistq, Smthwestern Medual School, Dalh, Texas
S. GAIT, Departmnt of Biorhemictry, Hebrew Universi+Hndassah Medical School,
Jerusalem. Israel
I. C. GUNSALUS, Depart- oJBiochemistry, Universaly of Illinois, Urbana, Illinair
H. A. 0.H ILL, Department oflnorganic Chemutry, Uniuerstty of Oxford, England
J. H. R. KAGI, Biochemical Institute. Universdy $Zurich, Switrerlnnd
B. G. MALMSTROM, Department of Bwchemistq, University OfGoteborg, Sweden
A. MEISTER, Department of Biochemistry, Cornell Medical Collpge, Npw York,N ew York
R. S. MELVILLE, National Instatute oJ General Medical Sciences, NIH, USPHS,B ethesda,
Maryland
M. OTTESEN, Carlsberg Laboratory. Copahagpn, Valby,D enmark
YC. A. OVCHINNIKOV, Shemyakin Institute for Chemisgr of Natural ProducLs, USSR
Academy of Sctares, Moscow, USSR
J. E. SCOTT,D epartment of Medical Biochemistry, Universtty of Manchester, England
E. C. SWTER, Laboratory of Biochemwtry,B . C. P. Jansen Institute, University of Amsterdam,
The Netherhuh
B. L. VALLEE, Biophysics Resfarch Laboratory, Department of Biologual Chemistry, Harvard
Medacd School, Boston, Mauuchusetls
K. YAGI, Institute uf Biochemutly, University of Nagoya Medical School,J apan
1
METHODS OF
BIOCHEMICAL ANALYSIS
Edited by DAVID GLICK
Cancer Biology Research Labwatcny
Stanford University Medical Center
Stunford, California
26
VOLUME
An Intemciencea Publication
- -
JOHN WILEY & SONS, New Chichester Brisbane Toronto
York
An lnterscience @Publication
Copyright 0 1980 by John Wiley & Sons, Inc.
All rights reserved. Published simultaneously in Canada.
Reproduction or translation of any part of this work
beyond that permitted by Sections 107 or 108 of the
1976 United States Copyright Act without the permission
of the copyright owner is unlawful. Requests for
permission or further information should be addressed
to the Permissions Department,J ohn Wiley & Sons, Inc.
Library of Congress Catalogue Card Number: 54-7232
ISBN 0-471-04758-8
Printed in the United States of America
1 0 9 8 7 6 3 4 3 2 1
METHODS OF BIOCHEMICAL ANALYSIS Volume 26
PREFACE
Annual review volumes dealing with many different fields of science
have proved their value repeatedly and are now widely used and well
established. These reviews have been concerned primarily with the results
of the developing fields, rather than with the techniques and methods
employed, and they have served to keep the ever-expanding scene within
the view of the investigator, the applier, the teacher, and the student.
It is particularly important that review services of this nature should
now be extended to cover methods and techniques, because it is becoming
increasingly difficult to keep abreast of the manifold experimental
innovations and improvements which constitute the limiting factor in
many cases for the growth of the experimental sciences. Concepts and
vision of creative scientists far outrun that which can actually be attained
in present practice. Therefore, an emphasis on methodology and
instrumentation is a fundamental need in order for material achievement
to keep in sight of the advance of useful ideas.
The volumes in this series are designed to try to meet the need in the
field of biochemical analysis. The topics to be included are chemical,
physical, microbiological, and if necessary, animal assays, as well as basic
techniques and instrumentation for the determination of enzymes,
vitamins, hormones, lipids, carbohydrates, proteins and their products,
minerals, antimetabolites, etc.
Certain chapters will deal with well-established methods or techniques
which have undergone sufficient improvement to merit recapitulation,
reappraisal, and new recommendations. Other chapters will be
concerned with essentially new approaches which bear promise of great
usefulness. Relatively few subjects can be included in any single volume,
but as they accumulate, these volumes should comprise a self-
modernizing encyclopedia of methods of biochemical analysis. By
judicious selection of topics it is planned that most subjects of current
importance will receive treatment in these volumes.
The general plan followed in the organization of the individual
chapters is a discussion of the background and previous work, a critical
V
vi PREFACE
evaluation of the various approaches, and a presentation of the
procedural details of the method or methods recommended by the
author. The presentation of the experimental details is to be given in a
manner that will furnish the laboratory worker with the complete
information required to carry out the analysis.
Within this comprehensive scheme the reader may note that the
treatments vary widely with respect to taste, style, and point of view. It is
the Editor’s policy to encourage individual expression in these
presentations because it is stifling to originality and justifiably annoying to
many authors to submerge themselves in a standard mold. Scientific
writing need not be as dull and uniform as it too often is. In certain
technical details, a consistent pattern is followed for the sake of
convenience, as in the form used for reference citations and indexing.
The success of the treatment of any topic will depend primarily on the
experience, critical ability, and capacity to communicate of the author.
Those invited to prepare the respective chapters are scientists who either
have originated the methods they discuss or have had intimate personal
experience with them.
It is the wish of the Advisory Board and the Editor to make this series of
volumes as useful as possible and to this end suggestions will be always
welcome.
DAVIDG LICK
METHODS OF BIOCHEMICAL ANALYSIS Volume 26
CONTENTS
The Use of the Avidin-Biotin Complex as a Tool in Molecular
Biology. By Edward A. Bayer and Meir Wilchek,
Department of Biophysics, The WeizmannI nstitute of
Science, Rehovot, Israel .................................... 1
Polarographic Measurement of Steady State Kinetics of Oxygen
Uptake by Biochemical Samples. By H. Degn, J. S.
Ludgaurd, and L. C. Peterson, Institute ofBiochemishy,
Odense University, O h e ,D enmark; A. Ormicki, Department
of Computer Sciences, Institute of Mathematics, Odense
University, Odense, Denmark. ............................. . 47
Analysis of Biological Macromolecules and Particles by Field-
Flow Fractionation. ByJ . Calvin Gdings,
Marcus N. Myers, Karin D. Caldwell, and
Susan R. Fisher, Department of Chemistry, University of Utah,
S a l t M City, Utah ....................................... 79
Recent Developments in the Stopped-Flow Method for the Study
of Fast Reactions. By Keitaro Hiromi, Laboratmy of Enzyme
Chemistry, Department of Food Science and Technology, Faculty
of Agriculture, Kyoto University, Kyoto,J apan ................ .,1 37
I
Peptide Mapping of Proteins. By Gordon T.J ames, Department of
Neurology, University of Colorado Medical Center, Denver,
Colorado ............................................. ,165
Solid-Phase Methods in Protein Sequence Analysis. By Richard A.
Luursen, Department of Chemistry, Boston Universzty,B oston,
Massachusetts; Werner Machleidt, Instimf ur Physwlogische
Chemk,P hysiRalishe Biochemie und Zellbiologie akr Universitat
Munchen, Munich ....................................... .201
vii
viii CONTENTS
Analysis of Cellular Electron Transport Systems in Liver
and Other Organs by Absorbance and Fluorescence
Techniques. By Helmut Sies and Bolko Brauser, Institutf ur
Physbbgisch Chemte, Physikalische Bwchemie und
Zellbiologie a!t?r Universitiit Munchenm, Munich ............... .285
High Temperature Gas-Liquid Chromatography in Lipid
Analysis. By V. A. VaverandA.N . Ushakov, Shemyakin
Institute of Bwmganu Chemistry, USSR Academy of Sciences,
Moscow ................................................ .32 7
AuthorIndex ................................................. 407
Subject Index ................................................ .417
Cumulative Author Index, Volumes 1-26 and Supplemental
Volume ............................................... .429
Cumulative Subject Index, Volumes 1-26 and Supplemental
Volume ...............................................
.440
Methods of Biochemical Analysis, b701ume 26
Edited by David Glick
Copyright 0 1980 by John Wiley & Sons, Inc.
METHODS OF BIOCHEMICAL ANALYSIS VOLUME 26
The of the Avidin-Biotin Complex
Use
a Tool in Molecular Biology
as
EDWARDA . BAYERan d MEIRW ILCHEK. Department ofBaophysics. The Weizmunn
Institute of Science. Rehwot. Israel
I . Introduction ...............................................
I1. Principle ...................................................
111. Preparation of Reactive Biotinyl Derivatives ............................... 5
1 . Amino Reagents ................................................... 7
2 . Carboxyl and Sugar Reagents ....................................... 7
3 . Thiol Reagents .................................................... 8
4 . Phenol and Imidazole Reagents ...................................... 8
IV . Assays for Avidin and Biotin ............................................ 9
v . Purification Studies: Affinity Chromatography .......................... 9
1 . Natural Biotin-Containing Systems ................................... 9
2 . "Tailor-Made" Biotin-Containing Systems ............................ 11
3. Experimental Procedures ........................................... 14
A . Preparation of Affinity Columns ............................... 14
a . Activation of Sepharose ................................. 14
b . Biotin Affinity Columns ................................ 14
Biocytin ............................................ 14
Biocytin Sepharose ................................... 14
Polymeric Biotin-Containing Columns .................. 14
c . Avidin Affinity Columns ................................ 15
Cyanogen Bromide Induced Immobilization ............ 15
Periodate-Induced Immobilization ..................... 15
Other Coupling Methods ............................. 15
B . Isolation Procedures ......................................... 15
VI . Locathation Studies: Affinity Cytochemistry ............................. 16
1. General Considerations ............................................. 16
2 . Localization of Functional Groups ................................... 17
A . Sugars ...................................................... 17
B . Amino Acid Residues .................... ............ 20
3 . Localization of Receptors ........................................... 22
4 . Other Systems ........................ ..................... 23
5 . Restrictions of the Method .......................................... 25
6 . Experimental Procedures ........................................... 26
A . Preparation of Avidin-Conjugated Markers ..................... 26
a . Fenitin-Avidin Conjugates via Reductive Alkylation ....... 26
1
EDWARDA . AND MEIRW ILCHEK
2 BAYER
b. GlutaraldehydeM ethod ....... ..................... 26
c. Other Coupling Methods .....
d. Other Markers ..............
e. Analysis of Conjugates ...............
B. Direct Biotinylationo f Functional Groups
a. Sialic Acid Residues .............
b. Galactose and N-Acetylgalactosa
C. Biotinylationo f Biologically Active Pro
D. Interaction Between Biotinylated Binding Protein and Receptor ... 29
VII. Miscellaneous Systems ............................
1. Phage Inactivation Studies ........
B. Experimental Procedures .......... ................ 29
2. Lymphocyte Stimulation ................. ...................... 31
3. Hormone-ReceptorI nteractions ..................................... 33
VIII. The Biotin Transport S m: An Affinity Labeling Study ................ 35
IX. Antibiotin Antibodies .............................................. 40
X. Conclusions ............................................... ... 41
Acknowledgments ....................... ....................... 42
References ...... ............................................... 42
I. INTRODUCTION
The high affinity constant between the glycoprotein avidin and the
vitamin biotin prompted early attention to the nature of this complex. To
obtain further insight into the properties of the avidin-biotin complex, in
the early 1950s Fraenkel-Conrat and co-workers ( 1952) purified avidin
and studied the effect of chemical modification on its activity. further
No
interest was taken in the complex until the end of the decade when Wakil
et al. (1958)a nd Lynen et al. (1959) discovered the coenzyme function of
covalently bound biotin. It became clear that avidin could be used as a tool
for characterizing biotin-requiring enzymes. In fact, the spatial position
of the avidin-bound biotin-containing subunit of transcarboxylase was
ultimately localized by high resolution electron microscopy (Green et al.,
1972; Green, 1972).
Sice 1963 Green has been the leading figure in the efforts to
understand this unique interaction by various biophysical and
biochemical methods (Green, 1975). However the innate reason for the
strong interaction between biotin and avidin is not yet known. Judging
from the structure of biotin (Figure l), it is difficult to understand why
such a simple molecule should such unprecedented affinity for
possess an
a given protein. Even more surprising, only the intact ureido ring is