THE EXPERIMENTAL MODELS OF FIRST
LIFE OBJECTS. THE ACTIONS OF
BIOLOGICAL ACTIVE SUBSTANCES TO
THESE MODELS.
1Yagolnik E.A., 2Alekseeva O.M.,
3Kim Yu. A.
1Tula State University, Tula,
2N.M.Emmanuel Institute of
Biochemical Physics, RAS,
Moscow,
3 Institute of Cell Biophysics,
Russian Academy of Sciences,
Pushchino, Moscow region
E-mails: yea_88@mail.ru;
olgavek@yandex;
yuk01@rambler.ru
Model phospholipids membranes imitated the main
2
components of bio-objects. We formed the
multilammelar liposomes with aid of hydration
of phospholipids thin films under the shacking
at warm phosphate buffer.
Biological active substances actions to these
experimental objects were tested.
Multilammelar liposomes that were artificially
formed from phospholipids bilayer membranes
were the adequate model of some intracellular
organelles.
• The liposomes are fairly interesting model for research of
primary bio-similar structures formation and action of different
materials on their organization, biologically active substances
(BAS). Among them were ions, simple non organic and organic
substances.
• Liposomes are organized due to the spontaneous process of
self-assembly of amphiphilic phospholipids molecules.
Hydrophobic and hydrophilic interactions stimulate the
liposomes formation with different structures in dependent of
hydration degree.
• Cell organelles are imitated by bilayer and multilayer
liposomes.
• Restructuring of cell compartments are simulated by means of
hexagonal and stack lipid structures.
• With the help of differential scanning microcalorimetry (DSC)
method we described any actions of some organic (Melafen
and phenozan and its derivatives) biologically active substance
on such model membranes. Multulammelar liposomes that
were formed from artificial individual phospholipid dimyristoilphosphatidylcholine (DMPC) were used for DSC
experiments. Liposomes were treated under the Melafen
presence from 10-17 M to-10-3 M).
3
Heat absorbtion
4
Thermo induced endothermic phase
transitions of phospholipids
(DSC) test.
Multilammelar liposomes
of DMPS
T
Pre - transition
Main transition
Lipid packing from more rankorder to free
Gel phase
Ripple-phase
Liquid crystal phase
speed of heat supplied (1 degree/min; 0,5 degree/min; 0,25 degree/min; 0,125 degree/min
one-eighth of degree)
to cells with control and experimental samples
Melafen influence to DMPC
thermo inducted parameters
– enthalpy, T, cooperatives
5
6
The structural effects of ICHFANs to the multulammelar DMPC liposomes
Redundant heat absorbtion
DMPC thermograms
control
ICHFANs
ICHFAN C16 formed the its own phase
under the concentrations 10-5, 10-4 М
at the belayer. This phase has another
thermo inductions parameters.
The domains of ICHFANs have the great
influencing to the rafts near the integral
and associated receptors and enzymes at
cellular membrane, by this its affect to the
functional activities.
ICHFANs С10 and С12
have the maximal effects, С8 and С16 have
the less effect, probably the length of
anchoring fatty acid residue С10, С12 is
the optimal.
Phenozan has not the fatty
acid tail; by this its influence is weaker.
Phenozan does not formed the own phase.
Its effects were occurred under the
biggest concentrations 10-4, 10-3 М
scheme of possible ways of liposomes
and cell interactions
Intracellular
vesicle formed
7
Next step in bio-structural
complicating may be the
building of hydrophobic peptide
molecules in such membranous
liposomes, or else grabbing
of hydrophilic ones to interior
space. Some regions of big
outer membranes may be
drowning, in interior space,
break away and so that form
small internal liposome with
mixed composition, as the
organelles predecessors.
The education of cell
compartments led on creating
of ionic and electrostatic
gradients that was able to
already emergence of some
functions.
Membrane fusion and vesicle introducing, than it were cat and form internal vesicles
8
• As of
experimental
object we used
the erythrocyte
ghost.
9
Melafen, ICHFAN-10, phenozan influence to the
thermo induced protein domains denaturation of
erythrocyte ghosts membranes suspension
phenozan
Erythrocyte ghosts thermograms
А- spektrin, actin; В1-ancyrin, demantin, Band
4,1, 4,2; В2- Band 3 cytoplasm fragment; СBand 3 – ion channels; D – membrane’s
blabbing
NH2
N
H2N
N
N
.
O
HOP(CH2OH)2
2 ICHFAN-10 10-6 М
control
ICHFAN-10 10-5
М
NH2
J /K
ethanol
Melafen
control
10
Conclusions:
• Certain small and large structural changes of
lipid and protein components of cell membrane
when biological active substances presence at
medium are being revealed by DSC test.
• So that, on first the making up stages of living
cells, some materials (as organic, and non
organic) changed the cellular structure and the
interaction of their components.