Ultracentrifugal Analysis in Theory and Experiment. A Conference Sponsored by the National Academy of Sciences with the Financial Support of the National Science Foundation; Held at the Rockefeller Institute from June 18 to June 21, 1962

279 Pages·1963·6.65 MB·English

by  J. W. Williams (Eds.)

ULTRACENTRIFUGAL
ANALYSIS
in Theory and Experiment
A Conference sponsored by the National Academy
of Sciences with the financial support of the
National Science Foundation) held at The Rockefeller
Institute from June 18 to June 21, 1962.
Edited by
J. W. Williams
Department of Chemistry
University of Wisconsin
Madison, Wisconsin
@
1963
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Participants in the Conference
Names in boldface type denote contributors to the volume. Numbers in parentheses
indicate the pages on which the authors' contributions begin.
E. T. Adams, Jr., Department of Chemistry, University of Wisconsin Madison, Wisconsin
f
(119)
W. J. Archibald, Department of Physics, Dalhousie University, Halifax, Nova Scotia,
Canada (37)
R. L. Baldwin, Department of Biochemistry, Stanford University School of Medicine,
Palo Alto, California (143)
J. W. BEAMS, Department of Physics, University of Virginia, Charlottesville, Virginia
C. 0. BECKMANN, Department of Chemistry, Columbia University, New York, New York
P. A. CHARLWOOD, National Institute for Medical Research, Mill Hill, London, N.W.7,
England
J. M. CREETH, Lister Institute, Chelsea Bridge Road, London, S.W. 1, England
H. G. ELIAS, Institut für chemische Technologie der Eidgen. Technischen Hochschule,
Zurich, Switzerland
S. ERLANDER, Northern Utilization Research and Development Division, United States
Department of Agriculture, Peoria, Illinois
Hiroshi Fujita, Department of Polymer Science, Osaka University, Osaka, Japan (119)
M. TH. GEHATIA, ASRCNP, Wright-Patterson Air Force Base, Dayton, Ohio
G. A. Gilbert, Department of Chemistry, The University, Birmingham, England (59, 73)
R. Haase, Institut für physikalische Chemie, Technische Hochschule, Aachen, Germany (13)
W. F. HARRINGTON, Mergenthaler Laboratory for Biology, The Johns Hopkins University,
Baltimore, Maryland
J. E. HEARST, Department of Chemistry, University of California, Berkeley, California
J. J. HERMANS, Chemstrand Research Center, Inc., Research Triangle Park, Durham,
North Carolina
G. J. Hooyman, Physics Laboratory, The University, Utrecht, The Netherlands (3)
R. C. LL. Jenkins, Department of Mathematical Physics, Portsmouth College of Technology
Portsmouth, England (59)
J. S. JOHNSON, Chemistry Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee
GERSON KEGELES, Department of Chemistry, Clark University, Worcester, Massachusetts
T. KOTAKA, Department of Biochemistry, Stanford University School of Medicine, Palo Alto,
California
AKIRA KOTERA, Department of Chemistry, Tokyo Kyoiku University, Otsuka, Tokyo, Japan
Stig Ljunggren, Division of Physical Chemistry, The Royal Institute of Technology,
Stockholm, Sweden (29)
V
vi PARTICIPANTS IN THE CONFERENCE
L. G. LONGSWORTH, The Rockefeller Institute, New York, New York
Victor J. MacCosham, Department of Chemistry, University of Wisconsin, Madison,
Wisconsin (249)
D. A. Machines, The Rockefeller Institute, New York, New York (xiii)
D. MCINTYRE, Polymer Structure Section, National Bureau of Standards, Washington,
D.C.
Matthew Meselson, Biological Laboratories, Harvard University, Cambridge, Massa-
chusetts (131)
Guenther Meyerhoff, Camille Dreyfus Laboratory, Research Triangle Institute, Durham,
North Carolina (47)
P. F. Mijnlieff, vanH Hoff Laboratory, The University, Utrecht, The Netherlands (81)
Girair M. Nazarian, Department of Chemistry, San Fernando State College, Northridge,
California (131)
J. B. NICHOLS, Experiment Station, du Pont de Nemours and Company, Wilmington,
Delaware
A. G. Ogston, Department of Physical Biochemistry, John Curtin School of Medical
Research, Australian National University, Canberra, Australia (263)
J. L. ONCLEY, Biophysics Research Division, Institute of Science and Technology, University
of Michigan, Ann Arbor, Michigan
LARS ONSAGER, Department of Chemistry, Yale University, New Haven, Connecticut
J. TH. G. OVERBEEK, van't Hoff Laboratory, The University, Utrecht, The Netherlands
K. O. PEDERSEN, Laboratory of Physical Chemistry, University of Uppsala, Uppsala,
Sweden
George Scatchard, Department of Chemistry, Massachusetts Institute of Technology,
Cambridge, Massachusetts (105)
H. K. Schachman, Biochemistry and Virus Laboratory, University of California, Berkeley,
California (171)
T. SHEDLOVSKY, The Rockefeller Institute, New York, New York
E. M. Shooter, Department of Biochemistry, Stanford University School of Medicine,
Palo Alto, California (143)
Rodes Trautman, Plum Island Animal Disease Laboratory, Animal Disease and Parasite
Research Division, Agricultural Research Service, United States Department of Agri-
culture, Greenport, Long Island, New York (203)
K. E. VAN HOLDE, Department of Chemistry, University of Illinois, Urbana, Illinois ^J
JEROME VINOGRAD, Church Laboratory, California Institute of Technology, Pasadena,
California
D. F. WAUGH, Department of Biophysics, Massachusetts Institute of Technology, Cam-
bridge, Massachusetts
J. W. Williams, Department of Chemistry, University of Wisconsin, Madison, Wisconsin
David A. Yphantis, The Rockefeller Institute, New York, New York (227)
The Aims of the Conference
Ultracentrifugal analysis may be said to have begun forty years ago.
Over this period of time there have been several organized meetings and
discussions to consider the subject in some or all of its phases. Of two such
affairs with U. S. National Academy of Sciences sponsorship, the first
"Conference on the Ultracentrifuge," with twenty participants, was held
at Shelter Island, New York, June 13-16, 1949. AJbrief account of the
topics discussed has been published. [Longsworth, Proc. Nail. Acad. Sei.
U. S. 36, 502 (1950)]. This report served a very useful purpose in that it
helped to bring to the attention of many workers in the field some of what
were then the outstanding problems in sedimentation analysis, and efforts
toward their elucidation were thereby stimulated. '
Now, more than a decade later, the subject has been tremendously ex-
panded both in the theory and practice of the methods themselves and in
their application to the solution of problems in biology and medicine. It
seemed again desirable to review and study the situation, for the ultra-
centrifuge is now known to provide a group of methods for the observation
and study of the redistribution of the components, especially in macro-
molecular systems, from which interesting conclusions may be drawn. As
in the earlier Conference, the attempt was made largely to avoid considera-
tion of the many problems connected with the applications to proteins,
polysaccharides, and polymers in general; rather it was planned to discuss
some of the outstanding mathematical and operational problems of the
ultracentrifuge, and to expand and enrich efforts toward finding their
solution.
This time it was inferred that a number of scientists—chemists, physi-
cists, and biologists alike—would like to see in print and in one place the
main papers and at least some indication of the directions taken by the
discussions which followed them. Such a record should constitute an invita-
tion to all to advance the knowledge of the sizes and configurations of any
number of biologically and technically important macromolecules as they
are influenced by the nature of the solvent medium. Again, it should bring
practicing biochemists and the more theoretical physical chemists closer
together. The attempt is here made to provide such an account.
Vll
viii AIMS OF THE CONFERENCE
Basic sedimentation theory now derives from the thermodynamics
either of equilibrium (sedimentation-diffusion equilibrium experiment) or
of irreversible processes (sedimentation transport experiment). At the time
of the earlier National Academy of Science Conference, kinetic theory was
used to derive the essential working equations for systems in transport,
but it is now recognized that such derivations were lacking both in rigor
and generality, though admittedly they did provide a "molecular picture''
of the process, a source of satisfaction to many investigators. The new
derivations are of great interest in that, for the simple two component
system, flows which occur in separate sedimentation and diffusion experi-
ments are formulated in terms of a single mobility. In the extension to
multicomponent systems now made possible, the restrictive nature of the
classical equations of Svedberg is clearly revealed. As the subject unfolds
it is tempting to remark that in ultracentrifugal analysis one deals with
processes which are seemingly simple in physical principle, but which are
at the same time very complicated in mathematical description. In a recent
monograph by one of the participants in this Conference, the scattered
fundamental theory of the subject (transport and equilibrium) has been
assembled and brought into a consistent framework. What now will become
evident is that there are several basic plans according to which transport
theory may be constructed, including the macro-dynamical approach of
Lamm.
Also, discussions are here presented of advances in describing the sedi-
mentation behavior in heterogenous solute systems, the effects of associa-
tion-dissociation processes, both in transport and at equilibrium, the
interactions in multicomponent systems, and the development of the sedi-
mentation equilibrium in a density gradient experiment.
In planning a meeting in which there could be informal yet productive
exchanges of ideas and mutual benefits it was felt to be necessary to limit
severely the number of participants. The basic criterion for selection was
subject matter; that portion of the program which had to do with the
scientific subjects to be considered was first set down. There then fell to
us the unhappy duty of making a somewhat arbitrary choice among a
number of investigators who have made significant contributions to certain
of the several subjects. It is regretted that more could not be accommodated.
The generous hospitality of President D. W. Bronk and Drs. L. G. Longs-
worth, T. Shedlovsky, and D. Yphantis, who made available and brought
into action the facilities of the Abby Aldrich Rockefeller Hall of The
Rockefeller Institute, was always available and greatly appreciated by all.
Finally, it is a pleasant task of the Editor:
(1) to express his own personal indebtedness to the members of The
AIMS OF THE CONFERENCE IX
Rockefeller Institute Staff cited above for their valuable advice and help
in the organization and operation of the Conference;
(2) to thank the officers of the National Academy of Sciences, in particu-
lar its President, Dr. D. W. Bronk, its Executive Officer, S. D. Cornell, and
its Business Manager, G. D. Meid, for their aid;
(3) to make grateful acknowledgment to the National Science Founda-
tion for a generous grant in support of the project;
(4) to express my appreciation to Academic Press for its willingness to
publish these Proceedings and the consideration with which it has acted;
(5) to acknowledge the aid of Dr. E. T. Adams, Jr., in seeing manuscripts
and proof-sheets to their ultimate end.
ADDENDUM
It had been our intention to ask to be printed a reasonably comprehensive
record of the discussions which followed the presentations of each of the
papers. The tape recordings of them left something to be desired. Also on
some few occasions the discussions were but obliquely related to the sub-
ject of the report which they followed. These situations, coupled with the
fact that several of the articles are rather long, have brought us to the
decision not to reproduce the comments, individual by individual. How-
ever, we have elected in a few cases to try to give a brief indication of the
paths into which such remarks were directed. If we have failed to under-
stand, the fault rests with us; all the participants did their part and did it
well.
We wish also to record a feeling of personal pleasure in noting the deep
interest of so many talented younger scientists. It was really evident in
their manuscripts, in their comments, and even most of all in their tête-à-
tete gatherings. This interest augurs well for the future of our subject;
with the courage of their years there will be another "Conference on the
Ultracentrifuge. ' '
J. W. WILLIAMS
Introductory Remarks
D. A. MaclNNES
Honorary Chairman
In the absence of President Detlev Bronk it is my pleasant task to wel-
come the participants of the Conference on the Ultracentrifuge at The
Rockefeller Institute.
A series of conferences on that instrument, of which this is the third,
have been held. The first of these was under the auspices of the New York
Academy of Sciences, and took place at the Museum of Natural History
on November 14 and 15, 1941. The papers presented at that conference
were collected into a monograph,1 which appears to have been of consid-
erable utility to workers of the field. The conference was one of a number
organized by the author and associates to provide a gathering of a group of
workers in limited fields to afford opportunity for close discussion of
matters of common interest. The great success of this first series of con-
ferences was, to an extent, their undoing, since they attracted larger and
larger attendances and, especially if the topics had general interest, became
little different from the meetings of the large scientific societies. Since it
did not appear feasible, under the organization of the New York Academy,
to limit the attendance at the conferences, the author, with the aid of Dr.
F. B. Jewett, then president of the National Academy of Sciences, started
a new series of conferences under the auspices of that society, in which
the participants could be limited to those who were competent to take
part in active discussion. It is argued, plausibly, that many more may get
profit from the discussions, but experience shows that, in general, the quality
of the discussion diminishes rapidly as the number of people present
increases.
The first conference in the new series was on ' 'Bioelectric Phenomena"
and was held at The Rockefeller Institute. Later ones took place at the
Rams Head Inn, on Shelter Island, New York. One of these was on the
1 Ann. N. Y. Acad. Sei. 18, 173-252 (1942).
xi
XÎi INTRODUCTORY REMARKS
ultracentrifuge and occurred on the three days, June 13 to 16, 1949. A
report on that conference was prepared by Dr. L. G. Longsworth and was
published in the Proceedings of the National Academy of Sciences?
The third conference in this series is now to be held here at The Rocke-
feller Institute. This conference differs from the two others in having inter-
national attendance, there being three participants each from The Nether-
lands, Japan, and England, two each from Sweden and Germany, and one
each from Australia, Canada, and Switzerland. W. J. Archibald, J. W.
Beams, L. G. Longsworth, D. A. Maclnnes, T. Shedlovsky, and J. W.
Williams were participants at all three of the conferences mentioned above.
The first two conferences certainly aided in the advance of the ultra-
centrifuge as an instrument for scientific research and it seems more than
likely that this third one will be equally or more useful.
2 Proc. Nail Acad. Set. U.S. 36, 502 (1950).
Thermodynamics of Diffusion and Sedimentation
G. J. HOOYMAN
Physics Laboratory, The University, Utrecht, The Netherlands
I. Introduction 3
II. General Theory 4
A. The Entropy Balance Equation 4
B. The Linear Laws and the Onsager Symmetry Relations 8
III. Applications 9
A. Binary Diffusion 9
B. Ternary Diffusion 9
C. Sedimentation Equilibrium 10
D. Velocity Sedimentation in Binary Systems 10
E. Velocity Sedimentation in Ternary Systems 11
F. Other Applications 11
References 12
I. Introduction
Since the previous "Conference on the Ultracentrifuge" in 1949 a com-
paratively new thermodynamic theory has been applied to transport
phenomena in the ultracentrifuge. This theory has been developed during
the last twenty years and goes by the name of thermodynamics of ir-
reversible processes or nonequilibrium thermodynamics. The basic ideas
were given as early as 1931 by Onsager (1931a, b). Although this extension
of ordinary thermodynamics is by now standard (de Groot and Mazur,
1962; Meixner and Reik, 1959) it might be useful to repeat the underlying
ideas and then give the application to diffusion and sedimentation (Hooy-
man et al, 1953; Hooyman, 1956a, b).
Classical thermodynamics is restricted to so-called reversible processes
and to equilibrium states and does not include the description of transport
phenomena. For these phenomena it only gives the second law in the
3