Low frequency dielectric

spectroscopy of biopolyelectrolytes

Institut za fiziku
Laboratory for biological physics
Silvia Tomi, Tomislav Vuleti,
Sanja Dolanski Babi,
Tomislav Ivek

Motivation

Proteins and DNA are organized moelcular machines

Physics of (self)organized molecular systems
Biomacromolecules are polyelectrolytes
Interactions and charge correlations in solution influence
the ordering and selforganization of biomacromolecules
Dielectric spectroscopy may provide information on the
structure and ordering of the ensemble, as well as on
conformation of single molecules

Tomislav Vuleti,
Institut za fiziku

Dielectric function

conductivity and
dielectric function

dD (t )

dt

D ( ) E ( )

t 0 E (t )

j 0 i E ( )

0 i

measured conductivity and dielectric

function
l
Y ________ Y G ( ) iC ( )
S
l C (
l G (
' ' ' __________
i
S
S

Tomislav Vuleti,
Institut za fiziku

Experimental setup
Precision impedance analyzer
Agilent 4294A: 40 Hz-110 MHz

Agilent BNC connectors

chamber

Temperature
control unit:
0 to 60C
Stability:
10 mK

Conductivity
chamber
For aqueous
samples:
1.5- 2000S/cm;
volume 50-200 L
Reproducibility 0.5%,
Long term (2 h) 2%

Pt

steel
casing

Tomislav Vuleti,
Institut za fiziku

Sample preparation and application

Genomic DNA fibers

100 L droplet

Closed chamber

DNA solution
Tomislav Vuleti,
Institut za fiziku

Measurements: complex conductivity

im
G()=Gexp()
Gbg()
C()=Cexp()
Cbg()

l G (

l C (
re
S
Tomislav Vuleti,
Institut za fiziku

Complex dielectric function ( ) ' ( ) i ' ' ( )

100

iste
otopine duge DNA
DNA vodene
in pure water
a1

25 oC

a2

prilagodba sume Cole Cole funkcija na

dobivenu kompleksnu dielektrinu funkciju

1
( )
1 (i )1 a

''

10
a4

NF
mod
103

100

104

a3

HF
105 106
(Hz)

107

108

vodene
otopine
dodanom soli
DNA in
1mMsNaCl
duge DNA

jakost relaksacijskog procesa

- srednje relaksacijsko vrijeme

1- distribucija relaks. vremena

b1

RESULT: two broad modes, whose position in

frequency and ampitude depend on the
concentration of the biopolyelectrolyte
and on the ionic strength of the solution

b2

10

b3
b4

LF
103

104

105 106
(Hz)

HF
107

108

Tomislav Vuleti,
Institut za fiziku

Fundamental scales of polyelectrolytes

LHF
LLF

L DNa
2

Einstein-Smoluchowski
equation

L- fundamental length scales

Tomislav Vuleti,
Institut za fiziku

Modern theoretical approaches

c* - crossover
concentration=
molecule mass /
swept volume

Semidilute regime

c* N mbp / N3a3
c* ~ 1 / Lc2 *

Dilute
regime

c*~N-2
Polydisperse genomic Na-DNA:
kbp
Contour length:
m
For the shortests fragments of 2 kbp:
0.006 mg/mL
Range of studied concentrations:

2 20
0.7 - 7
c*
0.01 - 5

Tomislav Vuleti,
Institut za fiziku

HF relaxation probes the structure

of the whole biopolymer solution Semidilute
regime
correlation length or mesh size
at crossover concentration:
c c* Lc = Na

Away from c* , should depend on c:

Lc (c* / c)m
solution property independent of N
N a(N-2 / c)m m=0.5

c-0.5

P.G.de Gennes et al.,J.Phys.(Paris),

37, 1461 (1976)
Tomislav Vuleti,
Institut za fiziku

HF mode length scale correlation length

Pure water

cDNA-0.33
cDNA-0.5

cDNA-0.5
deGennes
correlation length

cDNA
Hydrophobic polymers:

-0.33

Low DNA concentrations

No added salt Denaturation?
Locally fluctuating regions
With exposed hydrophopic cores
Tomislav Vuleti,
Institut za fiziku

HF mode length scale Debye screening

length
Added salt

-1

c-0.5

Added salt
screens
Coulomb
potential

Is= 1 mM added salt:

2Is > cDNA Debye length: -1=10nm

cDNA > 2Is corr.length:

Tomislav Vuleti,
Institut za fiziku

LF relaxation probes the structure

of the single biopolymer in solutionSemidilute

regime

Average size of the

chain
- chain is a random
walk
of correlation blobs
g monomers inside c-0.5 blob
g c 3
chain: N / g correlation blobs
chain size: R (N / g)0.5

R c-0.25

Tomislav Vuleti,
Institut za fiziku

LF mode length scale chain size

Pure water

LLF = R
cDNA-0.25

Tomislav Vuleti,
Institut za fiziku

LF mode length scale persistence

Added salt
length
Lp 50 nm
DNA structural
persistence length

50 nm

(r+l)
(r)

orientational
correlation of chain
segments
decreases as
<cos()>~exp(-l
/Lp)

Tomislav Vuleti,
Institut za fiziku

Conclusion

Two relaxation modes

Relaxation times are related to fundamental length
scales for (semi)dilute solutions
HF mode desribes the structure/organization of the
ensemble of macromolecules the solution structure
de Gennes correlation length
LF mode describes the conformation of a given molecule
Chain size
Adding salt promotes other length scales
Debye screeening length instead of the correlation length
Peristence length instead of the chain size
LFDS may provide more info on the polyelectrolyte then
X-rays
Tomislav Vuleti,
Institut za fiziku

Basic elements in the polyelectrolyte

theories
Poisson-Boltzmann equation

d 2 (r )
(r )

dr 2
0 r

Debye-Huckel approximation

zi e (r ) k BT

Linearized PB equation
Debyeva screening
length in a simple
electrolyte

d 2 (r )
2 (r )
2
dr

1 m

1 nm

0 r k BT
2e 2 N A

e r
(r )
r

1
I s mM

10
I s mM
Tomislav Vuleti,
Institut za fiziku