TIBS 14 - March 1989 89

Features
Talking Points
Deciphering the language of cells ecular mechanisms have received little
attention from the biochemistry com-
munity. Two popular textbooks of cel-
Tian Yow Tsong lular biochemistry, for instance, do not
include these essential activities in their
lists of methods of transcellular signal-
Long distance cell-to-cell or organism-to-organism communications may be ing. A prominent biochemist, in a
accomplished by transmission and reception of electromagnetic signals through recent conversation with the author,
membrane receptors or enzymes. Consistent with this idea is the observation that even labeled study of this type of cell-
membrane A T P a s e s a r e capable of absorbing energy from oscillating electric fields to-cell communication as 'astrology'
of defined frequency and amplitude and using it to perform chemical work. The and maintained that signals could only
concept of the 'electroconformational coupling' is used to explain how an electric be carried by 'the substance of chemis-
signal can modulate the activity of a membrane protein, and conversely, how an try', such as molecules or ions.
energy-dissipating reaction can produce an electric signal. Although any activity of a cell or an
organism must ultimately be account-
able or linked to reactions of mol-
Most biochemists agree that cells com- signals to warn or to inform other ecules, these reactions can be and most
municate with each other either by fishes. Multicellular signals and organ- likely are driven by physical forces.
direct exchange of metabolites through to-organ communications also use elec- Here we will consider how communica-
gap junctions, or by transfer of m e s s e n - tric impulses (see for example Ref. 6). tions through space by force fields
ger molecules or ions over distance. Again, direct transmittal of ligands are (electrical, magnetic, pressure, etc.,
Slime mold and yeast, for example, not essential. Birds, bats and micro- i.e. the substance of physics) may also
secrete cAMP to coordinate raovement organisms use radiowaves and mag- accomplish similar tasks and are uni-
of the cell population towards a center netic fields for orientation or to identify versally used by cells and organisms.
of aggregation at a certain stage of de- or determine the position of objects. The study of the molecular mechanisms
velopment. In eukaryotic cells neuro- All these activities are crucial to life of these modes of communication
transmitters, hormones, grow~:h factors, and are fascinating from a scientific might enable us to decipher the lan-
etc. regulate metabolism, growth, dif- viewpoint, but until recently their mol- guage of cells.
ferentiation, biosynthesis and loco-
motion. Thus, multicellular signals
originate from the interaction of 60 i i

ligands with membrane receptors of

target cells, which then activate a (a) (b)
¢-
cascade of biochemical reactions 1 3.
Communication by transmitting mol- o
4O
ecules or ions over space, however, is a
slow process and not effective over long
distances. Cells and organisms often
_ ~ • _ ~ ~ ~ _-
need rapid communications, and so
m~. 20
they have adopted other methods of 03

transcellular signaling. The n-Lost fami- o

E
liar of these is the use of sound waves:
birds sing to attract the opposite sex;
we speak to convey our thotLght. The 0 1 i i i i i i i i I

sound waves propagate through the 0 1 2 3 4 5 6 5 10 15 20 25 30 35

space. No messenger molecules or ions
are sent but the messages reach their
audience and are perceived. Another
familiar example is the electro-
communication of fishes 4'5. Certain
species of electric fishes are able to
send E O D (electric organ discharge)
7". Y. Tsong is at the Department o f Biochemistry,
University o f Minnesota College o f Biological
Sciences, St. Paul, MN55108, USA.
log (frequency in Hz) Stimulated voltage (V cm -~)
Fig. 1. Activation o f the Rb + (K+ )-pump o f Na +, K +-A TPase by an oscillating electric field, at 4°C (Ref.
19). (a) Human erythrocytes were exposed to an oscillating electric field o f 20 V cm -1 at different
frequencies for 1 h. Rb + uptake was monitored by the radioactive tracer, 8ORb+ (see text and Ref. 12).
Rb + uptake o f electric-field stimulated samples (0), stimulated samples treated with 0.2 mM ouabain
(D), non-stimulated samples (S), and non-stimulated sample treated with ouabain ( A ) are plotted
against the frequency o f the applied field. (b) The same experiment, with an electric field o f l k H z at
different field strengths. Symbols used are the s a m e a s in (a).
In similar experiments no ouabain-sensitive Rb + efflux was stimulated by the electric fields. The
cytoplasmic concentration o f Rb + w a s 27 raM, and the external concentration o f Rb + was 10 raM. Thus,
the field induced Rb + uptake was an active transport. No consumption o f A TP was detected.

© 1989, Elsevier Science Publishers Ltd, (UK) 0376-5067/89/$(12.l)0

90 T 1 B S 14 - M a r c h 1 9 8 9

(a) 200 mV to provide such energy.

Experimentally, high fields (in the
order of kV cm -l) have been used. In
P1 f
P2 these experiments either chloroplasts
were deprived of light energy or the
electron transport chain was inhibited.
The only possible source of energy was
that contained in the applied electric
Sin Sout field. These enzymes appear to be able
to absorb free energy from the external
electric fields to perform chemical
\ work. Signal transductions are by
P1S f P2S
nature similar to energy transductions.
Examples of corresponding processes
include energy absorption for signal
reception, energy conversion for signal
processing and energy transmission for
signal transmission.
Another type of experiment carried
---,%, - _ -~.~-- out with Na+,K+-ATPase is poten-
tially more informative19-22(D-S. Liu,
R. D. Astumian and T. Y. Tsong, sub-
mitted). Here the enzyme is shown to
utilize free energy transmitted through
an oscillating electric field to pump Na +
and K + against their respective concen-

-t-- tration gradients. Experiments with

red blood cells revealed three impor-
tant factors that determine the effi-
ciency of the transduction of electric
L_ energy. (1) The applied weak electric
field (20 V cm ~) must be amplified at
the site of the cell membrane to a
(b) (c) strength sufficient to shift the equilib-
rium of a chemical reaction. In this case
Fig. 2. A cyclic kinetic model which is shown to p u m p a neutral substrate against its concentration the field must be amplified by 1000
gradient when exposed to an oscillating electric field. (a) In the scheme, P2 is assumed to have a greater times to reach a transmembrane elec-
molar electric moment (AM) than P~. ['2 is" also assumed to have higher affinity for So,,,than PI has for tric field of 20 kV cm- i. A cell is like a
Sin. (b) When the oscillatingfield (indicated by sinusoidal curves) is in its positive phase, it induces a large
spherical shell made of insulating ma-
flux o f PI to P2 transition and a small flux o f PjS to P2S transition (both given in curved arrows). As the
result, there is a net clockwise flux o f enzyme conformational transitions, with a concomitant clockwise terial (lipid membrane) and according
flux orS. (c) When the electric field is"in the negative phase, it induces a large flux o f P2S to P IS transition to the Maxwell relation, an external
and a smaller flux o f P2 to P~ transition. The net result, again, is a clockwise flux o f enzyme electric field is amplified by approxi-
conformational transitions, with a concomitant clockwise flux o f S. The direction o f fluxes can be mately R J d times across the mem-
reversed if the affinity o f P I for Si~ is"greater than the affinity of P 2for So,,. brane (Ro and d being, respectively,
the outer radius of the cell and the
thickness of the membrane). Thus, the
Electric activation of membrane components must be large enough to cell membrane is a site of the field
proteins alter the activity of these molecules. amplification t2"13"23. (2) There is one
Recent experiments from several The use of high-intensity electric optimum frequency for activating the
laboratories have shown that many cel- fields that would produce a transmem- K+-pump (1 kHz) and another for acti-
lular functions are stimulated or sup- brane potential comparable to the vating the Na+-pump (1 MHz) 19'21
pressed by weak oscillating electro- endogenous potential of cells or organ- (Liu, Astumian and Tsong, submit-
magnetic fields. Pulsed electromag- elles provides another approach. Here ted). (3) There is also an optimum field
netic fields have been found to promote membrane enzymes are shown to strength (20 kV cm-I transmembrane
bond cell regeneration, DNA-, RNA- absorb energy from electric fields for electric field) for activating both
and protein biosyntheses7 10. The acti- doing chemical work (for reviews see pumpslg"21(Liu, Astumian and Tsong,
vity of many membrane enzymes are Refs 12 and 13). ATPases of chloro- submitted). Some results which show
also affected by weak electromagnetic plasts, thermophilic bacteria and mito- the frequency- and the field strength-
signals 1°J1 From a thermodynamic chondria have been induced to dependence of the Rb+-pumps arc
point of view, a weak electromagnetic synthesize ATP from ADP and Pi with given in Fig. 1.
field may influence a cellular function pulsed electric fields 14-18. ATP syn-
only if mechanisms exist which would thesis requires input of approximately Electroconformational coupling model
allow amplification of the signal. The 10 kcal (42 kJ) per mole of free energy; (ECC)
interaction energy between the ampli- an electric field must induce a trans- The above results on Na+',K +-
fied, effective field and certain cellular membrane potential of approximately ATPase imply that certain membrane
T I B S 14 - M a r c h 1 9 8 9 91

enzymes or receptors possess the abil- mit signals via the

ity to decipher electric signals of well same enzymes or
defined frequency and amplitude and
react accordingly. In the case of
receptors. The trans-
membrane electric
(~1)
Uu u U"'U -u

Na+,K+-ATPase and ATP synthe- potential of a cell

tases, free energy contained in the elec- does play a role in
tric fields is absorbed arid coupled transcellular com-
directly to drive an endergonic reac- munication. The
tion. This need not be always the case,
however. If a channel responds to an
electric signal, for example, it can
most familiar one is
the generation and
propagation of the
(b) 1
I,i I I
either open or close, thus modulating action potential in a
the local ion concentration (e.g. Ca 2+)
and electric potential. As a result, an
enzyme (or a number of enzymes) may
neuron. According
to the analysis based
on the ECC, model (C)
N U H0
be phosphorylated or dephosphory-
lated, causing changes in activity.
Many other mechanisms involving bio-
signals are expressed
only in oscillating, or
fluctuating force
Ill
chemical reactions are possible, as with fields I3"24-26. Figure
chemical signaling, but the primary 3 gives several types
process of recognition here', is electric of waveforms which (d)
signaling. have been shown to
The results of Na +,K+-ATPase also work with the ECC
allowed us to formulate a :mechanism model by the compu-
based on the concept of 'electroconfor- ter analyses 2~'. Elec-

mationai coupling' (ECC) for cellular
signal and energy transduct:ions 13'24'25.
This model postulates tha! a protein
can undergo conformational changes
by a coulombic interaction with an
oscillating electric field (or any oscillat-
ing force field with which lhe protein
can interact). When the frequency of
the electric field matches Lhe kinetic
characteristic of the conformational
transformation reaction, a phenom-
enological oscillation among different
conformers of the enzyme :s induced.
At the optimum field strength the con-
formers formed are functional. The
oscillation of these product:[ve confor-
mers when coupled to the binding of
ligands, such as K + and Na ~, can lead
to active pumping of the iigands (see
Fig. 2). Likewise, when the reaction is
coupled to the binding of A D P and Pi,
ATP can be synthesized, as in the case
of mitochondrial ATPase. The maxi-
mum energy transferred is AM-E, in
which AM is the difference in the molar
electric moment of the two conformers
and E is the field strength (AM is the
sum of changes in permanent and
induced dipole moments) For ex-
ample, the interaction energy for a con-
formational change involving a AM of
200 Debye under an electric field of 400
kV cm l (AhOof 200 mV) would be ~ 4
kcai (16.8 kJ) mol i.
In principle, each class of proteins is
adapted to respond to an oscillating
force field (electrical, sonic cr chemical
potential) of defined frequency and
strength. Conversely, chenfical reac-
tions of the reverse order would trans-
'°' U III
tric fields of these
waveforms have
been shown to drive
the enzyme modeled
in Fig. 2 to perform
Fig. 3. Various waveforms o f stochastic noise (a)-(c) and regularly
chemical work. Most oscillating (d) and (e) electric fields shown to drive the kinetic scheme o f
electrophysiological Fig. 2 f o r doing chemical work. The electric field must be sustained by
measurements re- an energy-dissipating process', such as the metabolite-supported electron
ported in the litera- transport or the light driven photo-oxidative reaction. In (a) both
ture, however, give amplitude and lifetime are random; in (b) lifetime is constant but
amplitude random; in (c) amplitude is constant but lifetime random; in
only constant values
(d) a cosine wave; and in (e) a square wave or meander function signal is
for the transmem-
shown. (See text and re)erences for detai&.)
brane potential of,
cells, including the resting potential of vicinity of an energy-transducing pro-
neurons. tein. A constant potential can easily be
modulated either by electron transport
Locally fluctuating transmembrane to become locally oscillatory or fluc-
electric fields tuating (as might be the case for the
This brings us to another question inner membrane in mitochondria), or
critical for the understanding of trans- by the opening and closing of an ion
cellular signaling. For an electrocon- channel with a characteristic frequency
formational change to occur, one need (as in neurons).
merely consider the short-range inter- There are other possible interpret-
action between a protein and an elec- ations of the above results on electric
tric field 13"e~e6. That is to say, the field activation of membrane ATPases.
time-average and space-average value In particular, the surface compartmen-
of transmembrane electric field is tal model of M. Blank 27, which is based
irrelevant in the analysis of the ECC on field-induced ion accumulation near
model. It is the local electric field that is the cell membrane surface, has been
pertinent. A coulombic interaction shown to reproduce some results
between two charges is inversely pro- obtained with the Na+,K+-ATPase.
portional to the square of the distance However, the ECC model is consistent
separating the two charges. The inter- with the electric property of most mem-
action energy fades away rather brane proteins 13"24-2~', and acknowl-
quickly. It is conceivable that the trans- edges the active role of some proteins.
membrane potential of a cell does Another advantage of the present
indeed display large amplitude fluctu- model is that it can bridge the gap
ation or oscillation when it is time- between the chemiosmotic hypothesis
resolved to microseconds or when it is and the conformational coupling hy-
recorded at a small region, say at the pothesis of the bioenergetics 2s 3~. Here
92
the electric p o t e n t i a l is a s s u m e d to be
r e s p o n s i b l e for the c o n f o r m a t i o n a l
c h a n g e o f A T P a s e . T h e a c t i v a t e d state
of the A T P a s e is t h e n c o u p l e d to d r i v e
an e n d e r g o n i c r e a c t i o n . L o c a l i z e d p r o -
ton m o v e m e n t is given the role of
m o d u l a t i n g the electric p o t e n t i a l to b e -
c o m e 'locally o s c i l l a t o r y ' , thus, allow-
ing the A T P a s e to t u r n o v e r 13"24-26"32.
A n e x p e r i m e n t a l c h a l l e n g e p o s e d by
the E C C m o d e l is to investigate the
p a t t e r n of the t r a n s m e m b r a n e electric
p o t e n t i a l of a cell o r an o r g a n e l l e in a
limited region. Is it s t a t i o n a r y o r locally
o s c i l l a t o r y ? This r e q u i r e s a time resolu-
tion of m a n y c h e m i c a l a n d physical
p a r a m e t e r s of a cell m e m b r a n e to
m i c r o s e c o n d s and a space r e s o l u t i o n to
nanometers. These parameters may
include c o n c e n t r a t i o n s o f Ca 2+, N a +,
K +, c A M P a n d h o r m o n e s , as well as
physical p r o p e r t i e s such as t r a n s m e m -
b r a n e electric field, o s m o t i c p r e s s u r e ,
local t h e r m a l e n e r g y a n d m e c h a n i c a l
force.
T e c h n i q u e s for fluorescence i m a g i n g
of t r a n s m e m b r a n e electric fields using
sensitive f l u o r e s c e n t dyes have r e c e n t l y
b e e n d e v e l o p e d 3~35. T h e m i c r o s e c o n d
r e s o l u t i o n of a m e m b r a n e p o t e n t i a l
i n d u c e d by a p u l s e d electric field is of
special i n t e r e s t to the p r e s e n t discus-
sion 35. A l t h o u g h space r e s o l u t i o n of
optical m i c r o s c o p y is l i m i t e d , f u t u r e
d e v e l o p m e n t s using m o r e a d v a n c e d
t e c h n o l o g i e s , might b e a b l e to i m p r o v e
u p o n it. If large a m p l i t u d e local fluctu-
ations o f a t r a n s m e m b r a n e electric field
is i n d e e d an i n h e r e n t f e a t u r e of a cell
m e m b r a n e , a n d if t h e y can be faithfully
r e c o r d e d , we might be able to a n a l y s e
the m e s s a g e s c o n t a i n e d in t h e s e sig-
nals. E v e n at the c u r r e n t state of instru-
m e n t a t i o n , the t i m e m a y be ripe for us
to l e a r n the l a n g u a g e of the cell at the
single cell level. H o p e f u l l y , we will
t h e n be able to c o n t r o l a n d r e g u l a t e cell
functions, such as m e t a b o l i s m , differ-
e n t i a t i o n a n d g r o w t h , by ' s p e a k i n g ' to
t h e m using the l a n g u a g e t h e y best
understand.
References
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Roberts, K. and Watson, J. D. (1983) in
Molecular Biology of the Cell, pp. 717-765.
Garland Publishing
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(1986) in Molecular Cell Biology, pp. 667-
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To transform or not to transform
To transform, or not to transform, that is the question.
Whether 'tis nobler in the cell to suffer
The ons and offs of multiple phosphorylations
Or to take up oncogenes against a sea of factors
And by mutating, change them. To mutate, to change,
No more, and by a change to say we end
The control, and the thousand second signals
The cell is heir to; 'tis a consummation
Devoutly to be wish'd. To mutate, to change
To change, perhaps to die; aT, there's the rub,
For in that transformation to malignancy what cancer may come
When we have altered genetic control
Must give us pause; there's the respect
That makes calamity of too much UV
For who would bear the protein kinase C,
The receptor's signal, the membrane channels or
Transcriptional control or AMP
The DAG and then the calcium flux
Which PIP does give occasion to,
When he himself might his mutation make
With a retrovirus? who would regulation hear
To be quiescent - such a weary life -
But that the dread of something aft,er change,
The unexplored mutation from who s alter~l state
No normal cell returns, puzzles the will
And makes us rather bear the controls we have
Than change to others that we know not of.
Thus regulation doth make cowards of us al~,
And thus the native hue of cell division
Is sicklied o'er with the pale cast of Go,
And cell divisions of great pitch and moment
With this regard their currents turn awry
And lose the name of mitosis.
J.L. CUTTS
Department of Medicine,
The University of New Mexico,
Albuquerque. NM 87131, USA.