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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

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Model phospholipids membranes imitated the main 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.

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  • 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).
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Thermo induced endothermic phase

transitions of phospholipids

Heat absorbtion

(DSC) test.

Multilammelar liposomes

of DMPS

T

Main transition

Pre - transition

Lipid packing from more rankorder to free

Gel phase

Ripple-phase

Liquid crystal phase

melafen influence to dmpc thermo inducted parameters enthalpy t cooperatives

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

5

Melafen influence to DMPC thermo inducted parameters – enthalpy, T, cooperatives
the structural effects of ichfans to the multulammelar dmpc liposomes

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The structural effects of ICHFANs to the multulammelar DMPC liposomes

ICHFAN C16 formed the its own phase

under the concentrations 10-5, 10-4М

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, 10-3М

DMPC thermograms

control

Redundant heat absorbtion

ICHFANs

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scheme of possible ways of liposomes

and cell interactions

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BAS

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.

Intracellular

vesicle formed

Membrane fusion and vesicle introducing, than it were cat and form internal vesicles

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  • As of experimental object we used the erythrocyte ghost.
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Melafen,ICHFAN-10,phenozan influence to the thermo induced protein domains denaturation of erythrocyte ghosts membranes suspension

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phenozan

Erythrocyte ghosts thermograms

А- spektrin, actin; В1-ancyrin, demantin, Band 4,1, 4,2; В2- Band 3 cytoplasm fragment; С- Band 3 – ion channels; D – membrane’s blabbing

control

2 ICHFAN-10 10-6 М

ICHFAN-1010-5 М

J /K

ethanol

Melafen

control

conclusions

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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.