Table Of ContentCELLULAR BIOENERGETICS: Role of Coupled Creatine Kinases
Developments in Molecular and Cellular Biochemistry
Series Editor: Naranjan S. Dhalla, Ph.D., FACC
1. V.A. Najjar (ed.): Biological Effects ofG lutamic Acid and Its Derivatives. 1981 ISBN 90-6193-841-4
2. V.A. Najjar (ed.): Immunologically Active Peptides. 1981 ISBN 90-6193-842-2
3. V.A. Najjar (ed.): Enzyme Induction and Modulation. 1983 ISBN 0-89838-583-0
4. V.A. Najjar and L. Lorand (eds.): Transglutaminase. 1984 ISBN 0-89838-593-8
5. G.J. van der Vusse (ed.): Lipid Metabolism in Normoxic and Ischemic Heart. 1989 ISBN 0-7923-0479-9
6. J.F.C. Glatz and GJ. van der Vusse (eds.): Cellular Fatty Acid-Binding Proteins. 1990
ISBN 0-7923-0896-4
7. H.E. Morgan (ed.): Molecular Mechanisms of Cellular Growth. 1991 ISBN 0-7923-1183-3
8. GJ. van der Vusse and H. Starn (eds.): Lipid Metabolism in the Healthy and Diseased Heart. 1992
ISBN 0-7923-1850-1
9. Y. Yazaki and S. Mochizuki (eds.): Cellular Function and Metabolism. 1993 ISBN 0-7923-2158-8
10. J.F.e. Glatz and GJ. van der Vusse (eds.): Cellular Fatty-Acid-Binding Proteins, II. 1993
ISBN 0-7923-2395-5
11. R.L. Khandelwal and J.H. Wang (eds.): Reversible Protein Phosphorylation in Cell Regulation. 1993
ISBN 0-7923-2637-7
12. J. Moss and P. Zahradka (eds.): ADP-Ribosylation: Metabolic Effects and Regulatory Functions. 1994
ISBN 0-7923-2951-1
13. V.A. Saks and R. Ventura-Clapier (eds.): Cellular Bioenergetics: Role of Coupled Creatine Kinases. 1994
ISBN 0-7923-2952-X
KLUWER ACADEMIC PUBLISHERS - DORDRECHT / BOSTON / LONDON
CELLULAR BIOENERGETICS:
Role of Coupled Creatine Kinases
edited by
v. A. SAKS
lAboratory ofB ioenergetics
Institute ofB ioenergetics
Institute of Chemical and Biological Physics
Ravala 10, Tallinn EE 0001, Estonia
Centre de Physiologie et Physiopathologie Cellulaires
Universite Joseph Fourier
2280, rue de la Piscine
F-38041 Grenoble Cedex, France
and
RENEE VENTURA-CLAPIER
Laboratoire de Cardiologie Cellulaire et Moleculaire
INSERM CJF 92-11
Universite de Paris-Sud, Faculte de Pharmacie
5, rue J~an:Baptiste Clement
F-9229(Chatenay-Malabry Cedex, France
Reprinted from Molecular and Cellular Biochemistry, Volumes 1331134 (1994)
KLUWER ACADEMIC PUBLISHERS
DORDRECHT I BOSTON I LONDON
Library of Congress Cataloging-In-Publication Data
Cellular bioenergetics: role of coupled creatine kinases / edited by
V.A. Saks and Renee Ventura-Clapier.
p. cm. --(Developments in molecular and cellular biochemistry: 13)
"Reprinted from Molecular and cellular biochemistry."
ISBN 0-7923-2952-X (acid-free paper)
1. Creatine kinase. 2. Energy metabolism. I. Saks, V.A.
II. Ventura-Clapier, Renee. III. Molecular and cellular biochemistry. IV. Series:
Developments in molecular and cellular biochemistry: v. 13.
QP606.073045 1994
574.87'6 --dc20
ISBN 0-7923-2952-X
Published by Kluwer Academic Publishers,
P.O. Box 17, 3300 AA Dordrecht, The Netherlands.
Kluwer Academic Publishers incorporates
the publishing programmes of
D. Reidel, Martinus Nijhoff, Dr W. Junk and MTP Press.
Sold and distributed in the U.S.A. and Canada
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© 1994 Kluwer Academic Publishers
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CONTENTS
Preface
I - MUSCLE ENERGY METABOLISM
la. V.A Saks and R. Ventura-Clapier: Introduction: history of the problem 5
1b . H.M. Eppenberger: A brief summary of the history of the detection of creatine kinase isoenzymes 9
2. M.D. Osbakken: Metabolic regulation of in vivo myocardial contractile function: multiparameter analysis 13
3. Y. Ishida, I. Riesinger, T. Wallimann and RJ. Paul: Compartmentation of ATP synthesis and utilization in smooth muscle: roles
of aerobic glycolysis and creatine kinase 39
4. M. Wyss and T. Wallimann: Creatine metabolism and the consequences of creatine depletion in muscle 51
11-SUBSTRATE AND CREATINE KINASE ISOENZYME COMPARTMENTATION
1. D. Brdiczka and T. Wallimann: The importance of the outer mitochondrial compartment in regulation of energy metabolism 69
2. F.N. Gellerich, M. Kapischke, W. Kunz, W. Neumann, A. Kuznetsov, D. Brdiczka and K. Nicolay: The influence of the cytosolic
oncotic pressure on the permeability of the mitochondrial outer membrane for ADP: implications for the kinetic properties of
mitochondrial creatine kinase and for ADP channelling into the intermembrane space 85
3. S. Soboll, A Conrad and S. Hebisch: Influence of mitochondrial creatine kinase on the mitochondriaVextramitochondrial
distribution of high energy phosphates in muscle tissue: evidence for a leak in the creatine shuttle 105
4. T. Schnyder, M. Rojo, R Furter and T. Wallimann: The structure of mitochondrial creatine kinase and its membrane binding
properties 115
5. R Ventura-Clapier, V. Veksler and lA. Hoerter: Myofibrillar creatine kinase and cardiac contraction 125
6. F. Savabi: Interaction of creatine kinase and adenylate systems in muscle cells 145
III - CREATINE KINASES AND METABOLIC INTEGRATION
1. V.A Saks, Z.A Khuchua, E.V. Vasilyeva, O. Yu. Belikova and AV. Kuznetsov: Metabolic compartmentation and substrate
channelling in muscle cells. Role of coupled creatine kinases in in vivo regulation of cellular respiration - a synthesis 155
2. T. Wallimann and W. Hemmer: Creatine kinase in non-muscle tissues and cells 193
3. IF. Clark: The creatine kinase system in smooth muscle 221
IV - MOLECULAR BIOLOGY OF CREATINE KINASES
1. RM. Payne and AW. Strauss: Expression of the mitochondrial creatine kinase genes 235
2. S.M. Miihlebach, M. Gross, T. Wirz, T. Wallimann, l-C. Perriard and M. Wyss: Sequence homology and structure predictions of
the creatine kinase isoenzymes 245
3. 1 van Deursen and B. Wieringa: Approaching the multifaceted nature of energy metabolism: inactivation of the cytosolic creatine
kinases via homologous recombination in mouse embryonic stem cells 263
V - DEVELOPMENT AND PATHOLOGICAL ALTERATIONS OF CREATINE KINASES
1. lA Hoerter, R. Ventura-Clapier and A Kuznetsov: Compartmentation of creatine kinases during perinatal development of
mammalian heart 277
2. V. Veksler and R. Ventura-Clapier: In situ study of myofibrils, mitochondria and bound creatine kinases in experimental
cardiomyopathies 287
3. E.K. Seppet and V.A Saks: Thyroid hormones and the creatine kinase system in cardiac cells 299
VI - METABOLIC REGULATION: THEORETICAL BASIS
1. B.N. Kholodenko, M. Cascante and H.V. Westerhoff: Control theory of metabolic channelling 313
2. M.K. Aliev and V.A Saks: Mathematical modeling of intracellular transport processes and the creatine kinase systems: a
probability approach 333
Molecular and Cellular Biochemistry 133/l34: 1, 1994.
© 1994 Kluwer Academic Publishers.
Preface
The first book in the English language all about one single enzyme - creatine kinase - was published, to our knowledge,
in 1980, and that was Heart Creatine Kinase, edited by William E. Jacobus and Joanne S. Ingwall. Since that time, the
progress of studies of creatine kinase systems in different cells has been very impressive. There have been several
publications covering some part of the topic, such as Myocardial and Skeletal Muscle Bioenergetics, edited by N. Braut
bar (Plenum Press, 1986), a monograph by S. Lyslova and V. Stephanov Phospagen Kinases (CRC Press, 1991) and
several review articles. However, the main part of information on the functional role, structure and molecular biology
of creatine kinase is scattered in several hundreds of publications. The intention of the editors of this volume was to
invite all authors who are most active at the present time in the experimental research of creatine kinases to summarize
their work and make the information of the 'state of the art' easily available to a wider scientific audience, especially to
young investigators who are just now entering this field of research. Since the functional role of coupled creatine
kinases is directly related to the phenomenon of compartmentation and structural organization of metabolic networks,
we also invited experts in these related areas to contribute to this volume. Almost all invited authors responded enthu
siastically and the results of this collective work are presented here for the attention of the interested reader. We hope
that the volume will be of interest for many biochemists, biophysicists, physiologists as well as medical research work
ers.
The work of the editors in the preparation of this volume was possible due to the generous help from 'Institut de la
Sante et de la Recherche Medicale', France. Permanent support from Prof. N. Dhalla, President of the International
Society for Heart Research, is gratefully recognized.
Valdur A. Saks and Renee Ventura-Clapier
Laboratoire de Cardiologie Cellulaire et Moleculaire
INSERM CJF 92-11
Universite de Paris-Sud, Faculte de Pharmacie
5, rue Jean-Baptiste Clement
F-92296 Chatenay-Malabry Cedex, France
PART I
MUSCLE ENERGY METABOLISM
Molecular and Cellular Biochemistry 133/134: 5-8, 1994.
© 1994 Kluwer Academic Publishers.
'My work is not a piece of
writing designed to meet
the taste of an immediate public,
1-1 Introduction:
but was done to last forever'
Thucydides, 460-400 Be, Athens
history of the problem
'The Peloponnesian War'
Rather controversial and interesting history of phos who have traced back in the evolutionary time-scale the
phocreatine (PCr) research is already well described in moment when mi-CK appeared - a very long time ago,
several books and reviews [1-5]. In considering the de and surely not for nothing. Why it happened became
velopment of ideas of physiological role of PCr in mus clear, however, only after 1964. Again, S. Bessman in
cle, we may see movement of scientific opinions from cooperation with Fonyo [14] was first experimentally to
one extreme position to another. Sometimes it looks like demonstrate its functional role by showing that creatine
movement of a pendulum. After works of Lundsgaard is able to exert acceptor control of respiration. New in
which ended the 'lactate theory' of contraction, there dependent line of support for the central role of phos
was a 'phosphagen' era, when phosphocreatine was con phocreatine and creatine kinase system was found in
sidered as a direct source of energy for contraction [1, 2, studies of metabolically inhibited or ischemic hearts by
6]. Subsequent discoveries of creatine kinase reaction by Gercken and Schlette and Gudbjarnason and coworkers
Lohmann [7], ATPase reaction of myosin by Engelhardt [15,16], when contraction was shown to stop at high cel
and Lyubimova [8] and direct demonstration of utiliza lular levels of ATP, but in parallel to the tissue level of
tion of ATP in muscular contraction by Davies group [9, PCr. These two works attracted attention to the phe
10], however, reduced PCr to the role of simple energy nomenon of ATP compartmentation in the cells as a ba
buffer, a source for extra ATP supplied, if necessary, via sis for the important role of creatine kinase system. This
equilibrium creatine kinase reaction in cytoplasm at in concept - compartmentation of adenine nucleotides and
creased workloads. This is a point of view which still oc other metabolites - is now well verified (see chapters 1-3
cupies the minds of many learned men and can be found and 111-1) and most obviously will serve as a basis for our
in very recent publications. However, already at the understanding of cellular bioenergetics and integrated
time of important works by Davies [1, 9, 10], there were cellular metabolic systems.
investigators who began considering alternative points After discovery of mi-CK and compartmentation of
of views. One of those pioneers who started pushing the adenine nucleotides, it became clear that for real under
PCr pendulum back again was Samuel Bessman. At that standing of the physiological role of PCr in the cells, de
time, his ideas were mostly based on good intuition and tailed biochemical information of the behavior of CK
logical consideration of the mechanisms of acceptor isoenzymes in cellular structures was necessary.
control of respiration and they led him to formulation of From the very beginning, and up to our days, an im
the concept of phosphocreatine shuttle [11, 12, see 4, 5 portant role in these studies was played by the Switzer
for reviews). Very slowly, the experimental material also land group of Eppenberger, Wallimann, Perriard and
started to accumulate in favour of such more active role their coworkers. Their work is reviewed comprehen
of phosphocreatine, and the pendulum of opinions start sively and they are active authors of this volume (chap
ed slowly moving again from the shadow, energy reserve ters 1-3, 1-4, 11-1, 11-4, 111-2, IV-2) with impressive pre
area, in direction of the central role of PCr in muscle sentation of new data including presentation of the pri
cells. A decisive event was the discovery of mitochon mary structures of 26 guanidino kinases (chapter IV-2).
drial form of creatine kinase by Jacobs, Heldt and Klin Important contribution into these studies has been
genberg in 1964 [13]. In this issue, you can find many pa made by William E. Jacobus who, in cooperation with
pers solely devoted to this isoenzyme of creatine kinase Albert Lehninger, published in 1973 a paper on mi-CK
which is encoded by two separate genes different from which is still very well read as a fine phenomenological
those for M or B type creatine kinases (chapter IV-1). description of control of mitochondrial respiration by
The central role of this creatine kinase isoenzyme is em mitochondrial creatine kinase [17]. Equally important
phasized by Fig. 3 in chapter IV-2 by Miihlebach et al. was the contribution by another American group - Ing-
6
wall and Bittl, who have performed important series of tienzyme systems. This volume contains an important
energy flux measurements by NMR magnetization sat work by Kholodenko, Cascante and Westerhoff (chap
uration transfer technique. Some contribution into the ter VI-I) which leads the reader into the field of these
field of creatine kinase research has been made in our ideas. This new approach contains a novel and potential
laboratories by applying physiological methods to show ly powerful theory of metabolic control in organized sys
the importance of myofibrillar creatine kinase for con tems which may be much more efficient than that in di
trol of muscular contraction (chapter 11-5) and in studies luted solutions and may lead us to a better understand
of the mechanisms offunctional coupling of creatine ki ing of muscle biochemistry and biochemical regulation.
nases with other cellular systems (reviewed in chapter Another example of theoretical treatment of substrate
III-I). It is not possible to list here the names of all in channelling is given in chapter VI-2.
vestigators who have worked for finding the solution of In the experimental part of this volume there is a
the problems, but whose contribution is recognized in group of papers from three different laboratories consis
the lists of references. The results of all these works are tently describing the rather new and potentially very im
summarized by the Scheme 1, which shows connection portant phenomenon for metabolic regulation - the low
of creatine kinase to the systems of energy production permeability of the mitochondrial outer membrane for
(mitochondria) and energy utilization (myofibrils and adenine nucleotides and its possible role in regulation of
cellular membranes) in cardiac cells. Because of their cellularrespiration in cells in vivo (chapters II-I, II- 2 and
specific localization and interaction with other cellular III- 1). This is a very new and unexpected group of re
systems by a mechanism of direct substrate channelling sults, but a suspicion that the data collected in the ex
in multienzyme complexes, these creatine kinase isoen periments with mitochondria in vitro are not very help
zymes integrate energy metabolism by forming efficient ful for explaining the in vivo phenomena because of the
phosphocreatine pathway (shuttle) [4,5], or circuit (see loss of some important factors during isolation has been
chapter 111-2) for intracellular energy transport. Several clearly expressed in recent publications and the papers
versions of this Scheme will appear in other chapters of collected in this volume show that such a suspicion was
this volume (chapters 111-1 and 111-2) emphasizing in de highly justified. If confirmed in further studies, these re
tails its different aspects. sults may open new perspectives for studies of cellular
This scheme is only one of the good examples of the respiration in vivo.
importance of compartmentation, both of enzymes and Chapter IV-3 of this volume contains description of a
substrates in the cells. In literature, we can see rapidly new technology of creatine kinase gene manipulation.
increasing number of publications on compartmenta Even if we take into account very significant adaptive
tion of different substances including even cations, in mechanisms which may compensate for the switching
the cells (see chapter 111-1 for review). There is no doubt off ofthe creatine kinase gene and change the initial me
that this development will strengthen in the future and tabolic situation (see chapters 1-4 and III- 2), this is a very
reflects the complexity of intracellular reality. In this promising new area of research. Gene structure and ex
volume, classical approaches and methods for studies of pression for creatine kinase isoenzymes have been stud
muscle energetics are summarized by M. Osbakken ied in details and are described in this volume by Payne
(chapter 1-2), and as emphasized by the author, no clear and Strauss (chapter IV-I) and Miihlebach et al. (chap
answer was found for intracellular regulatory mecha ter IV-2). These studies are important for understanding
nisms of energy metabolism. What seems now impor of the developmental changes and pathological alter
tant to do is not only to use new techniques of research ation of the creatine kinase systems (chapters V-I, V-2,
but first of all to account for the complex nature of the V-3).
intracellular medium with characteristic compartmen Very interestingly, detailed studies of smooth muscle
tation phenomena, multienzyme organized structures showed high degree of compartmentation of metabolic
and direct interaction between their components - sub systems in these cells (chapter 1-3) in spite of rather low
strate - product channeling. In other words, it is prob rates of energy turnover, and the presence of creatine
ably time now to reject the idea of intracellular medium kinases which seem to be actively involved in the energy
being a simple aqueous solution as an historical necessi transport (chapters 1-3 and III- 3). Thus, it is clear that, in
ty but outlived simplification. Instead, it is time to in many tissues, the creatine kinases play more important
corporate into the metabolic research the very serious role than that of a simple energy buffer. In muscle and
achievements of physical chemistry of organized mul- brain mitochondria, the coupled creatine kinase reac-
7
*
*
* *
* * * * * * * * ** * * * *
* *
* * * * * * * * *
*
glucose *
* * * * * *
fr .. fatty acids * *
* K Na
*
* ....
Glycoly"sis .
IT
Cr ..... +
f +
...... PCr -r
.
" ~
"
" "
" "
£ .. .... ..
PCr"~ ~ ~~
• '.. . *
~
" ""PC,······
* *
PCr "" \Cr
"'.~
I mllD.Sic
'I'
actin ••
*
*
• MgATP
*
Calcium
&.
Creatine kinase
tension
Fig. 1. Excitation-contraction coupling in adult mammalian cardiac cells and creatine kinase pathway. Calcium entry into the cell is triggered by
action potential and activates contractile proteins. Creatine kinase isoenzymes are bound to sites of energy production (inner mitochondrial mem
brane and glycolytic clusters in cytosol) and to sites of energy utilization (myofibrils, sarcoplasmic reticulum, plasma membrane) in close association
with ATPases. SR, sarcoplasmic reticulum; IT, T tubules; lea, calcium current.
tion seems to function as an effective feedback signal atine kinases in the cells, and find a steady central posi
amplifier (chapter 111-1). However, its functional inter tion for the pendulum of opinions.
action with the adenylate kinase system as well as with Taken together, all these works give not only compre
hexokinase should be taken into account in explaining hensive account for the results of the research of cre
the cellular mechanisms of metabolic regulation by sub atine kinase systems, but also formulate new concepts
strate channelling via different specialized pathways concerning the structural and functional organization of
(chapters 11-1, 11-2 and 11-6). In this way we may ap integrated metabolic systems and their regulation in the
proach more realistic understanding of the role of cre- cell. We hope that many readers will find the experimen-
