[Study of P-glycoprotein effect on the lipid monolayer properties by the Langmuir-Blodgett technique]. / Issledovanie metodom Lengmiura-Blodzhett vliianiia P-glikoproteina na svoistva lipidnykh monosloev.

Expression of the P-glycoprotein (Pgp) is proved to be one of the main reasons for the development of the multidrug resistance (MDR) phenotype by cancer cells. The effect of Pgp on the properties of lipid monolayers was studied using membrane fractions of sensitive and Pgp over-expressing multidrug resistance cancer cells containing 11, 24 or 32% of Pgp relative to the total content of membrane proteins. The effect of the Pgp membrane concentration on the properties of monolayers prepared from the membrane fractions was analyzed by the Langmuir-Blodgett method. The subphase composition was found to play a critical role in the stability of monolayers at any Pgp concentration. The optimal subphase comprised 10 mM tris-HCl buffer, pH 6.5, which made it possible to create very stable monolayer films with the pressure of collapse of about 30-40 mN/m. Monolayers prepared from membrane fractions of sensitive cells and cells containing the maximum (32%) amount of Pgp were found to be much more stable compared with fractions comprising 11 or 24% of Pgp. The analysis of monolayer compression dynamics revealed three distinct stages: (1) the self-organization of lipid molecules, which is characterized by an abrupt change of surface potential; (2) the compression of Pgp molecules at the constant potential of monolayers; and (3) the compression of lipid molecules, which is characterized by a quasilinear increase of both pressure and surface potential. It was shown that the specific surface areas of monolayers formed from sensitive and Pgp-enriched membranes containing 11 or 24% of Pgp are very similar, whereas the surface area of the monolayer formed from membranes containing 32% of Pgp is nearly 1.5-fold greater. This fact may reflect the effect of the threshold rearrangement of the structure of lipid molecules or cooperative modifications of lipid-Pgp interactions induced by the increase in the Pgp content from 24 to 32%. The effect of verapamil, a well-known Pgp modulator, on the properties of monolayers was studied. It was show that verapamil is able to induce changes of the surface of Pgp-containing monolayers, and these modifications are maximal at the Pgp:verapamil 1:1 molar ratio. The data present the first experimental evidence for the possible intervention of Pgp modulator into the processes of lipid-lipid or lipid-Pgp cooperative interactions within Pgp-enriched membranes.

[Synthesis of amphiphilic photochromic benzo-15(18)-crown-5(6)-ethers and their properties in monolayers]. / Sintez amfifil'nykh fotokhromnykh benzo-15(18)-kraun-5(6)-éfirov i ikh issledovaniie v monosloiakh.

New amphiphilic photochromic benzo-15(18)-crown-5(6) ethers (APC) differing in the position of the octadecyl substituent and the size of the crown cavity were synthesized. The compounds form stable monolayers in the air/water and air/alkaline metal salt solution interfaces. The results of the pressure isotherm measurements, atomic force microscopy (AFM), and electronic spectroscopy show that the structure of the monolayers formed depends on the structure of the parent APC and the nature of the cation in salt solutions. The area per molecule of APC in the monolayer (specific area) is the smallest on the water surface and increases by 20-40% on the aqueous subphase surface with an increasing concentration of salts therein to indicate the formation of APC complexes with the metal cations. When the hydrophobic aliphatic substituent is displaced from position 3 to position 5 of the benzothiazole ring, the specific area on the surface of water and subphases decreases twofold, which indicates the compactization of the monolayer on this modification. A reversible E-Z-photoisomerization of APC was found in the monolayers formed in the salt solution/air interface. The features of the reaction are defined by the specific organization of the amphiphilic molecules in the monolayer and by the nature of the cation.

[The study of hydrolysis of new lipid-like substrates and trilaurin in monolayers catalyzed with the lipase from Pseudomonas fluorescens]. / Issledovanie gidroliza v monosloiakh novykh lipidopodobnykh substratov i trilaurina pod deistviem lipazy Pseudomonas fluorescens.

A simple method for determining the enzymic hydrolysis parameters of lipid-like substrates and trilaurin assembled in monolayers at the water-air interface was suggested. At a surface pressure of 10 mN/m, the initial rates of lipolysis were found to be proportional to the decrease in area of the substrate monolayer caused by the enzymic hydrolysis in a single-compartment Langmuir balance. The kinetic parameters for the hydrolysis of trilaurin and three 1,3-dilaurylpseudoglycerides acetylated in position 2 with an amino acid (phenylalanine, leucine, or valine) catalyzed with lipase from Pseudomonas fluorescens were determined. Unlike models of enzymic hydrolysis that neglect the thickness of the substrate monolayer, our method allows the determination of kinetic parameters in standard dimensions. The values of kcat for the synthetic pseudoglycerides were found to be significantly higher than that for trilaurin, while the values of Km(app) were close. This may be due to the presence of positively charged primary amino groups in the molecules of pseudoglycerides.

[Molecular design of polymeric materials for biotechnology and medicine]. / Molekuliarnoe konstruirovanie polimernykh materialov dlia biotekhnologii i meditsiny.

This review describes new polymer materials for biomedical applications developed in the Polymers for Biology Laboratory of the Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences. These include composite rigid sorbents for biochromatography, polymer dispersions for immunoassay, polymer hydrogels for immobilization of enzymes and cells, and polymer ultra thin films as biomembrane models and materials for biosensors. Some general and specific properties of these new materials and models as well as examples of their applications are discussed.

[Effect of surfactants on peroxidase activity. II. Effect of cationic surfactants]. / Vliianie poverkhnostno-aktivnykh veshchestv na aktivnost' peroxidazy. II. Vliianie kationnykh PAV.

The effect of various cationic surfactants on the initial rate of the 5-aminosalicylic acid peroxidation with horseradish peroxidase was studied. With increasing concentration of the surfactant, the rate first increased and then decreased. We attributed these changes to a primary activation of the enzyme and/or an increase in the concentration of the substrate in the region of the active site and to the subsequent inactivation of the protein with surfactant followed by its denaturing, respectively.

[Cholesterol in the plasmalemma of endotheliocytes in the normal aorta and in hypercholesterolemia (scanning electron microscopy research)]. / Kholesterin v plazmolemme éndoteliotsitov aorty v norme i pri giperkholesterinemii (skaniruiushchee élektronno-mikroskopicheskoe issledovanie).

Using a specific latex marker (SLM) and scanning electron microscopy, distribution of cholesterin in plasmolemma of endotheliocytes of the thoracic part of the rabbit aorta has been studied under normal conditions and at experimental atherosclerosis, as well as on the surface of liposomes and flat bilayer films. SLM reacts with a high specificity directly with cholesterin of the membrane. When studying microtopography of cholesterin, it is possible to distinguish microdomains with various density of distribution of SLM particles. Amount of SLM per 1 mcm2 of the artificial membrane surfaces grows at increase of the cholesterin-phospholipid (Ch/Phl) index of the membrane; Ch/Phl can bind with the membranes beginning only with the threshold value of the Ch/Phl index equal to 0.5. At hypercholesterolemia amount of SLM on the luminal surface of plasmolemma of the aortal endotheliocytes increases by 78%. Hence, at hypercholesterolemia in the luminal part of plasmolemma of the aortal endotheliocytes the Ch/Phl index increases; this should be taken into consideration, when observing mechanisms of reorganization of the endothelial cells layer at atherosclerosis.

[Polymer monolayers with immobilized bacteriorhodopsin]. / Polimernye monosloi s immobilizovannym bakteriorodopsinom.

Monolayers of the membrane photosensitive protein, bacteriorhodopsin, have been prepared and investigated. The protein is able to reversibly change areas of monolayers in the course of photoactivation. Chemical immobilization of bacteriorhodopsin in the polymer lipid layer allows limit the protein's conformational mobility in the excited state of photocyle, which is important for study of molecular mechanisms of photoreception and design of biosensor devices.

[Synthesis of polymerizable phospholipid derivatives and creation of polymer monolayers and liposomes with immobilized bacteriorhodopsin]. / Sintez polimerizuemykh proizvodnykh fosfolipidov i sozdanie polimernykh monosloev i liposom na ikh osnove s immobilizovannym bakteriorodopsinom.

Polymerizable phospholipid derivatives with N-acryloyl groups have been synthesised; monomeric and polymeric monolayers and liposomes based on these compounds have been prepared and investigated. Bacteriorhodopsin immobilized in the monomeric and polymeric liposomes was shown to retain its native activity.