METMBS

'00 lnternalional

Conference

J.A. ThszyIiski**J.A.

Brown and B. Trpisova

Department of Physics University of Alberta Edmonton, Alberta, T6G 2Jl Canada

Abstract

The possibility

of signalling

via microdipole

tubtde filaments moment. tubtdin

of the cytoskeleton

is investigated

using two different The first dimer

models based on tubulin's

model involves dipole flips in the either spontaThe second We find fluctuations in field guidance. polarization. that thermal

which may propagate

neously or by electric model of signalling main walls this latter of dielectric

studied is based on moving do-

case, however,

significantly disturb the propagation of such solitary waves unless strong electric fields are present.

microtubules, biological waves, polarization

signalling,

solitary

Introd

uction

Signalling by various means is necessary in order to regulate the complex behavior of living systems. The use of signaling molecules which are first packaged, then dumped outside the cell where they spread diffusively facilitates cellular communication.[I] Synaptic signalling is a refined version of paracine signalling in which neurotransmitter molecules are released from nerve cell's synapses.[2] Endocrine signalling USes hormones which are reverted into the cirsystem where they travel over long Within the cell, the mechanisms signal transduction are less clear. The cyhas been implicated in intracellular by many researchers. Electromagsignalling has also been involved and its
00
also at: Starlab NV /SA, Boulevard St. Michel 1040, Brussels, Belgium.

advantage is great speed of propagation. [3] Microtubules (MT's) are cytoskeletal protein filaments which have been shown to respond to both electric and magnetic fields aligning themselves to field lines [4]. Each of the MT's subunits, dimmers tubulin, has an of electric dipole moment that contributes to the overall polarity of the structure. The magnitude of this dipole moment p has been estimated as the product of an elementary charge e with a spacing of 8nm corresponding to the size of the dimmer.This gives a value of Po = 1.28.10-27Cm (=384 debye) which is very close to the value obtained from structural calculations based on the tubulin crystallography [5]. The latter gives 337 debye for the axial protofilament component of the tubulin dipole moment [6]. Our earlier Monte Carlo simulation of the dipole-dipole interactions between the dimmers a microtubule demonstrated that of such a lattice of dipoles may be ordered at or below physiological temperatures [7]. The ordering can be either ferroelectric (for the socalled 13A lattice) or antiferroelectric (for the 13B lattice) It is tantalizing that it might be this net polarization of the MT lattice which is responsible for guiding the motor proteins such as kinesin and dynein which travel in opposite directions along the MT .

Dipole Flips and via Microtubules

Signalling

We propose that the orientation of the individual dipoles may be flipped due to a confor-

mational change of the tubulin dimmer resulting from a GTP hydrolysis or from physical interactions (e.g. dipole-dipole forces) [8]. We then study the propagation of signals in the form of dipole patterns on the background of an ordered lattice of a ferroelectric or antiferroelectric type [9]. We have used Monte Carlo simulations [10] to model the problem (see Fig. 1). Our model was inspired by Hameroff et al [11] who developed a cellular automaton approach whereby a discrete charge on tubulin was able to hop. In our model, a dipole moment was instead assigned to each dimmer whose orientation was allowed to change. The interaction energy between two neighboring dipoles is [121

along the MT unless some additional mechanism is added. Since there is nothing to direct the propagation of the defect, it takes a random walk about the MT and its energy is gradually dissipated to the rest of the MT. The efficient propagation of signals could be restored by (i) the application of an external field which would bias signal propagation; (ii) an asymmetry in the dipole structure which simply makes it more favour able to propagate in a particular direction; or (iii) mechanical stress if dipoles are coupled to a lattice distortion. The third mechanism would be the result of a piezo-electric effect which was claimed to be a property of MT's a long time ago.

PI

.p2

-3(PI r3

.ii)(P2

.ii)

Eint
~

(I)
138 138
14A 14A

where ~ is the relative permittivity of the medium, ~o is the permittivity of free space, ~ is the ",th tubulin dimer's electric dipole moment. n is the normal vector pointing from the position of the first dipole to the second dipole and r is the distance separating the dipoles. It is known that MT's are assembled from GTP-rich tubulin dimmers and that this GTP is hydrolyzed rapidly after the addition of the tubulin subunit. What is not yet known is what happens to this energy. We are proposing that some of the energy is stored in the lattice through a conformational change of the protein dimer. The hydrolysis of GTP releases about 4.6 kcal/mol of energy which the lattice could use to flip conformational states of individual tubulin dimmers, each flip would require up to 2.0 kcal/mol of energy [9]. The energy might then propagate along the MT through a sequenceof dipole flips as the lattice reorients to accommodate the additional energy. These conformational changes or flips are believed to be the result of a mobile electron. It may be localized at one of two binding sites in the tubuliD molecule. Movement of the electron from one binding site to the other causes the tubulin dimmer and its electric dipole to re-orient. Unlike the Hameroff model [111,we do not observe the smooth propagation of signals

(a)

(b)

(c)

(d)

Figure I. Portions of different microtubule lattices with dipole order illustrated. Light boxes represent the dipole "up" state while dark boxes represent the dipole "down" state. (a) The I3B lattice above its critical temperature is characterized by dipole disorder, (b) the I3B lattice below its critical temperature is antiferroelectrically ordered; (c) The I4A lattice is ferroelectrically ordered below the critical temperature (d) but it also supports domain wall patterns (e) The I4A lattice just below the critical temperaure also supports large defect structures. Notice that smaller thermal defects dot the structure.

METMBS

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The existence of an ordered phase is crucial if the MT is to be able to process information. When the lattice is not organized, this signifies that thermal fluctuations dominate over the dipole-dipole interactions and that entropy dominates the lattice. Upon such a background, any signals introduced to the lattice rapidly vanish (thermalize). We have also been able to demonstrate that the application of a large axial field of 106V/m along the MT causes nearly all dipoles to orient themselves in the direction which most closely follows that of the field. Thus, a wave of dipole flips is induced along the MT as the field is translated along the MT. This is similar to what happens as an action potential moves along an axon. The large field is felt by MTs in the vicinity of the cell membrane. Suppose the field is oriented in a direction which favours an alternate ordering for the lattice, such as is the case in MT, these dipoles will re-orient themselves. The field acts like a pump and stores energy in the lattice of dimmers. Once the field has passed, these dimmers may return to their original configuration and release their stored energy. A weaker field does not actually create defects by changing the orientation of dipoles in the ground state, but can act as a bias and direct the movement of any existing defects. Our simulations place a firm limit on the strength of the dipoles required for selforganization. The required dipole strength of about I x 10-21 C.m is comparable but slightly larger than the value which we estimated for tubulin. If the angle between the dipole directubulin's conformational states than predicted, an even larger dipole be required. Should the dipole than this, informaprocessing via dipole flips must be ruled

tric phase of MT dipoles. We also believe that dipole flips are coupled linearly to a conformational charge in the MT lattice due to MT's piezoelectricity [16]. Based on the above assumptions, it has been demonstrated elsewhere that the Hamiltonian associated with the dynamics of a dipole placed at site n in a MT protofilament with N tubulin dimmers can be represented by the following equation [13]

; l, I

Propagation main Wails

of Dipolar

Do-

In eq. (2) the variable Un represents the projection on the protofilament axis of the distortion of the dimmer which is in the {3 state with respect to the Q state, 4M(dun/dt)2 is the kinetic energy of the tubulin molecule of mass M, 4K(Un+l -Un)2 represents the elastic energy that originates from the restoring elastic force$ characterized by a constant K that act between each two dimmers. The quartic doublewell potential energy V(Un) = -~u~ + Tu~ approximates the average effect of the surrounding dipoles on the dipole at site n. V(Un) can be viewed as a Landau n.ee energy expansion where Un corresponds to the order parameter and the coefficients Q2 and Q4 are characteristics of the physical system. Assuming that the phase transition from a ferroelectric to a paraelectric phase in a MT is a secondorder phase transition, Q4 is a positive constant and Q2 = a2(Tc -T), where a2 > 0. The coefficient c represents a bias field (electric or pressure gradient) acting on the dynamic variable uno Also, Ro is the equilibrium distance between two neighbouring tubulin dimmers. Based on the Hamiltonian in eq. (2) we obtain the following equation of motion in the continuum limit and with the addition of a friction term proportional to 1

seen in our simulations above that walls occur naturally in the ferroelec-

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The values of the constants in eq. (3) can be determined as follows [13): The mass of the tubulin dimmer was calculated as M = 1.83 x 10-22kg. The constants K and Ro are related to the velocity of longitudinal sound waves through the formula K m, where Vo is the sound velocity. Due to lack of direct measurement in MT's we take the velocity of sound in DNA, Vo = 1700 ms-l [14), to apply for MT's. The coefficients a2 = a2(Tc -T) and a4 for a MT are also not known. However, inorganic crystals exist in which ferrodistortive domain walls can be formed. For example, for the crystal PbsGe3Gll below the critical temperature, a4 = 1.6 x 1024Jm-4 and a2 = 10Jm-2 K-t. If the MT is in the ferroelectric phase then the critical temperature Tc can be approximately taken as 350K and T is body temperature 310K. This gives for the coefficient a2 ~ 400Jm-2. To estimate the damping coefficient 'Y, the tubulin dimmer can be considered a sphere of radius R = 4 x 10-9m and mass M that is moving in a fluid of viscosity 1]. Assuming that a MT is mainly surrounded by water molecules, 1] can be taken as the viscosity of water. At body temperature, 1] = l]water = 6.9 X 10-4kgm-ls-l. Consequently, 'Y = 61rRl] = 5.2 x 10-llkgs-l. For a constant electric field E, eq. (3) can be solved analytically [13) and takes the form of a wave that travels at a constant velocity v [10), with a moving coordinate introduced as follows f. = x -vt)

and

t/I(f.) = ~, Uo

Uo = (~)1/2. Q4

(7)

The travelling kink wave solution of eq. (5) is listed in (151. The solution that corresponds to the kink wave moving with the velocity v > O is (see fig. 2)

) 1/J1-1/J2 1/J(f. = 1/J2-r 1 + ~("'I-1/12)/h'

where 1/J1and 1/J2satisfy the cubic equation

(8)

(1/1

1/Jl)(1/J

1/J2)(1/J

1/J3)

1/1"-1/1-

a. (9)

It can be shown that the velocity of propagation of this domain wall structure follows an approximate formula given by
v ~ ~(~)1/2qE (10)

c=
~

la21

-V2)

1/2

(X

vt)

'4

The partial differential equation (3) then reduces to an ordinary differential equation lfl;'!/J ~+P~ where dtt
1/J3 + 1/J + q

(5)

p= [Ma2(V~

V'Y

-V2)]1/2'

~(~)1/2 Q2 Q2

(6)

'YO.2 2 The maximum electric field along the MT has been estimated at Emax ~ 2 .6 106 V /m which gives the corresponding kink velocity of v ~ 1 .2 m/s. The velocity v depends linearly on the electric field E. This means that larger electric fields on MTs will generate kinks moving at larger velocities and vice versa. The magnitude of the intrinsic electric field of a MT can be altered if the MT is subjected to an external electric field generated, for example, by a membrane or by other MTs. If the external electric field is oriented in the same direction as the intrinsic field of the MT, this will result in a faster propagation of kinks. If the external and intrinsic electric fields are oriented in the opposite direction, the propagation of kinks will be slowed down or reoriented when the magnitude of the external field is larger than the magnitude of the intrinsic field of the MT. In the previous section it was shown that sufficiently strong external fields can alter the direction in which the dipoles are aligned when the MT is in the ferroelectric phase. Then the external field and the intrinsic MT field would be oriented in the same direction. This would change the

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direction of propagation and increase the velocity of the kink waves. When the total electric field to which the dipoles on the tubulin dimmers subjected is zero the kinks will not are propagate at all. According to the approximation of the charge distribution in a MT chosen here the magnitude of the intrinsic field of a MT also depends on its length. This means that MTs with different sizesmay support kink waves travelling at different velocities. We have also investigated the effects of imPurity potentials on the propagation of kinks in the MT lattice. Both potential wells and bumps slow down moving domain walls due to the force generated by their gradients. A critical amplitude of the potential was found at which the kink is stopped completely. Its value was determined numerically to be 10-21 J which is on the order of thermal noise at physiological temperatures. This result indicates that under more realistic conditions of temperature, electric fields and pressures, the propagation of signals in the form of dipolar domain walls in a stochastic process strongly dependent on thermal fluctuations and hence the model presented in the first part of the paper is quite realistic.

Acknow

ledgements

This work was supported by grants from NSERC and the Consciousness Studies program administered by the University of Arizona.

References [1] B. Alberts, D. Bray, J. Lewis, M. Raff, K. RA:>berts, J.D. Watson, Molecular biology of and the cell, Garland Publishing, London, 1994. [2] M. V. Volkentstein, General Biophysics, Academic Press, New York, 1983. [31 G. Albrecht-Buehler, The Centrosome, chapter Speculation about the function and formation of centrioles and basal bodies. , pages 69-102, Academic Press, San Diego, 1992. [4] P.M. Vassilev, R.T. Dronzine, M.P. Vassileva, and G.A. Georgiev, Parallel Arrays of Microtubules Formed in Electric and Magnetic Fields, Bioscience Reports 2, p. 1025-1029 (1982). [5] E. Nogales, S.G. Wolf and K.H. Downing, Structure of the Alpha-Beta Thbulin dimmer by Electron Crystallography, Nature 39, 199 (1998). [61 J.A. Brown, Ph.D. thesis, Universityof Alberta, Edmonton, 1999. [71quad J .A. Thszynski, S. Hameroff, M. V. Sataric, B. Thpisova, and M.L.A. Nip, Ferroelectric behavior in microtubule dipole lattices: Implications for information processing, signaling and assembly/disassembly, J. Theor. BioI. 174,371-380 (1995). [81 B. Thpisova and J .A. Thszynski, Phys. Rev. E 55, 3288 (1997). [91 J.A. Brown and J.A. Thszynski, Phys. Rev. E 56, 5834 (1997). [11 K. Binder, Monte Carlo simulation in statistical physics: an introduction, SpringerVerlag, New York, 1988. [111 S. Hameroff, S.A. Smith and R.C. Watt, Automaton model of dynamic organization in microtubules, Ann. N.Y. Acad. Sci. 466, 949952 (1986).

c ~ 0
00.6

t t t t

t t t t

06.07

..c..o7

07

0.2.-01

-.~7

x(mj

Figure 2. A domain wall between two subchains of a MT protofilament in which the tubulin dirners are in two different states.

{121 J.D. Jackson, ClassicalElectrodynamics, John Wiley and Sons, Toronto, 1975. {131 M. V. Sataric, R.B. Zakula, J .A. Thszynski, A model of the energy transfer mechanism in microtubules involving domain-walltype 597 solitons, (1993). Physical Review E 48, p. 589-

{141 M.B. Hakim, S.M. Lindsay and J. powell, The speed of sound in DNA, Biopolymers 23, p. 1185-1192 (1984). {151 J.M. Dixon, J.A. Thszynski and M. Otwinowski, Special analytical solution of the damped-anharmonic-oscillator equation, Physical Review A 44, p. 3484-3491 (1991). {161 H. Athenstaedt, Pyroelectric and Piezoelectric properties of Vertebrates, Ann. N. Y. Acad. Sci. 238, 68--94(1974).