Argon-water Interface Research Articles

Influence of gas aggregation on water-solid interface: molecular simulation

ABSTRACT In this study, we conducted a series of molecular dynamics simulations of gas accumulation on homogeneous graphite substrates with a focus on the structure of fluid in the region near the substrate. The fluid density distribution revealed that epitaxial layers and gas enrichment layer arise from the adsorption of fluid particles on solids. Our analysis suggests that interfacial gas molecules transform into a liquid-like state and water molecules tend to cluster within the thin region adjacent to the substrate as the strength of gas–solid interaction increases. We then calculated the argon-water interfacial tension using three different methods: the Young-Laplace equation, direct integration method, and force balance method. Our analysis suggests that higher pressure within the gaseous domain tends to reduce tension at the curved argon-water interface. Although widely adopted to describe tension at gas–liquid interfaces, our results indicate that Bakker's equation is not necessarily suitable for the calculation of tension at fluid-solid interfaces. Our results indicate that fluid-solid interfacial properties and the morphology of the gaseous domain are influenced by the liquid-like epitaxial gas layer and gas enrichment layer, which could be attributed to the highly energetic nature of gas molecules and the tendency of gasses to adsorb on solids.

New Route for the Production of Thermosensitive Polymer With Pulsed Arc Discharge at the Argon–Water Interface

Poly-N-isopropylacrylamide (PNIPAM) is well known about an exhibition of coil-to-globule transition below the lower critical solution temperature (32°C) in aqueous media. Because of this temperature near the human body, it has focused on as one of the functional polymers available to industries. However, the synthesis of PNIPAM has been conventionally conducted in combined harmful solvents for considerable long operating times. In this paper, we aimed to develop a novel technique that PNIPAM can be produced in a rapid and selective manner with pulsed discharges at ambient temperature and pressure. Reactions were carried out using a stainless steel batch-type reactor (inner volume: 15 mL) in the presence of 2,2'-Azobis (2-methylpropionamidine) dihydrochloride as an initiator at 25°C-30°C, 0.89 M NIPAM aqueous solution, and pulse repetition rates 1-50 pps for 1-15-min treatment times. As the results, in most cases, PNIPAMs were able to be produced within a couple of minutes in relative high production yield and selectivity.

Menthol-induced action potentials in Conocephalum conicum as a result of unspecific interactions between menthol and the lipid phase of the plasma membrane.

Our previous study has shown that the liverwort Conocephalum conicum generates action potentials (APs) in response to both temperature drop and menthol, which are also activators of the TRPM8 (transient receptor potential melastatin 8) receptor in animals. Not only similarities but also differences between electrical reactions to menthol and cooling observed in the liverwort aroused our interest in the action of menthol at the molecular level. Patch-clamp investigations have shown that menthol causes a reduction of current flowing through slow vacuolar (SV) channels to 29 ± 10% of the initial value (n = 9); simultaneously, it does not influence magnitudes of currents passing through a single SV channel. This may point to an unspecific interaction between menthol and the lipid phase of the membrane. An influence of menthol on lipid organization in membranes was investigated in two-component monomolecular layers formed with menthol and dipalmitoylphosphatidylcholine (DPPC) at the argon-water interface. Analyses of the mean molecular area parameters vs the molar fraction of the menthol component have shown over-additivity (approximately 20 Å(2) ) in the region of high molar fractions of menthol. Infrared absorption spectroscopy studies have shown that menthol, most probably, induces breaking of a hydrogen bond network formed by ester carbonyl groups and water bridges in the lipid membrane and binds to the polar head group region of DPPC. We conclude that the disruption in the lipid phase of the membrane influences ion channels and/or pumps and subsequently causes generation of APs in excitable plants such as C. conicum.

Supramolecular Organization of the Main Photosynthetic Antenna Complex LHCII: A Monomolecular Layer Study

The light-harvesting pigment-protein complex LHCII is a main antenna complex of the photosynthetic apparatus of plants, responsible for collecting light energy and also for photoprotection against overexcitation-induced damage. Realization of both functions depends on molecular organization of the complex. Monolayer technique has been applied to address the problem of supramolecular organization of LHCII. Analysis of the isotherms of compression of monomolecular films formed at the argon-water interface shows that LHCII appears in two phases: one characterized by the specific molecular area characteristic of trimeric and one of monomeric organization of LHCII. Monolayers of LHCII were deposited by means of the Langmuir-Blodgett technique to solid supports and examined by means of AFM, FTIR, fluorescence spectroscopy, and fluorescence lifetime imaging microscopy (FLIM). FTIR analysis shows that organization of the trimers of LHCII within a monolayer is associated with formation of intermolecular hydrogen bonds between neighboring polypeptides. The linear-dichroism FTIR analysis reveals that polypeptide fragments involved in intermolecular interactions are oriented at an angle of 67 degrees with respect to the normal axis to the plane of the layer. Fluorescence and fluorescence lifetime analysis reveal that the organization of LHCII within monolayers is associated with formation of the low-lying excitonic energy levels that can be potentially responsible for excess excitation quenching. FLIM and AFM reveal heterogeneous organization of LHCII monolayers, in particular, formation of ring-like structures. The potential of LHCII to form molecular structures characterized by pigment excitonic interactions is discussed in terms of regulation of the photosynthetic accessory function and photoprotection against overexcitation-induced damage.

Organization of polyene antibiotic amphotericin B at the argon–water interface

Amphotericin B (AmB) is a life-saving antibiotic, used to treat deep-seated mycotic infections. Both the therapeutic and toxic side effects of AmB are directly dependent on its molecular organization. Organization of AmB was studied in monocomponent monomolecular layers formed at the argon–water interface, by means of polarized and non-polarized electronic absorption spectroscopy and analyzed in terms of the exciton splitting theory. The results provide direct indication that AmB forms spontaneously dimers that can be assembled into molecular structures characterized by homogeneous orientational distribution in the monolayer, interpreted as cylindrical pores. The structures are not stable at surface pressures higher than 20 mN/m and therefore dimers are concluded as abundant molecular organization forms of AmB in biomembranes. Possibility of stabilization of the cylindrical structures, at higher surface pressures, by other molecules, e.g. sterols, is also discussed.

Secondary structure and orientation of the pore-forming toxin lysenin in a sphingomyelin-containing membrane

Lysenin is a sphingomyelin-recognizing toxin which forms stable oligomers upon membrane binding and causes cell lysis. To get insight into the mechanism of the transition of lysenin from a soluble to a membrane-bound form, surface activity of the protein and its binding to lipid membranes were studied using tensiometric measurements, Fourier-transform infrared spectroscopy (FTIR) and FTIR-linear dichroism. The results showed cooperative adsorption of recombinant lysenin-His at the argon–water interface from the water subphase which suggested self-association of lysenin-His in solution. An assembly of premature oligomers by lysenin-His in solution was confirmed by blue native gel electrophoresis. When a monolayer composed of sphingomyelin and cholesterol was present at the interface, the rate of insertion of lysenin-His into the monolayer was considerably enhanced. Analysis of FTIR spectra of soluble lysenin-His demonstrated that the protein contained 27% β-sheet, 28% aggregated β-strands, 10% α-helix, 23% turns and loops and 12% different kinds of aggregated forms. In membrane-bound lysenin-His the total content of α-helices, turns and loops, and β-structures did not change, however, the 1636cm −1 β-sheet band increased from 18% to 31% at the expense of the 1680cm −1 β-sheet structure. Spectral analysis of the amide I band showed that the α-helical component was oriented with at 41° to the normal to the membrane, indicating that this protein segment could be anchored in the hydrophobic core of the membrane.

New BODIPY lipid probes for fluorescence studies of membranes

Many fluorescent lipid probes tend to loop back to the membrane interface when attached to a lipid acyl chain rather than embedding deeply into the bilayer. To achieve maximum embedding of BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) fluorophore into the bilayer apolar region, a series of sn-2 acyl-labeled phosphatidylcholines was synthesized bearing 4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene-8-yl (Me(4)-BODIPY-8) at the end of C(3)-, C(5)-, C(7)-, or C(9)-acyl. A strategy was used of symmetrically dispersing the methyl groups at BODIPY ring positions 1, 3, 5, and 7 to decrease fluorophore polarity. Iodide quenching of the phosphatidylcholine probes in bilayer vesicles confirmed that the Me(4)-BODIPY-8 fluorophore was embedded in the bilayer. Parallax analysis of Me(4)-BODIPY-8 fluorescence quenching by phosphatidylcholines containing iodide at different positions along the sn-2 acyl chain indicated that the penetration depth of Me(4)-BODIPY-8 into the bilayer was determined by the length of the linking acyl chain. Evaluation using monolayers showed minimal perturbation of <10 mol% probe in fluid-phase and cholesterol-enriched phosphatidylcholine. Spectral characterization in monolayers and bilayers confirmed the retention of many features of other BODIPY derivatives (i.e., absorption and emission wavelength maxima near 498 nm and approximately 506-515 nm) but also showed the absence of the 620-630 nm peak associated with BODIPY dimer fluorescence and the presence of a 570 nm emission shoulder at high Me(4)-BODIPY-8 surface concentrations. We conclude that the new probes should have versatile utility in membrane studies, especially when precise location of the reporter group is needed.

Interaction of isomeric forms of xanthophyll pigment zeaxanthin with dipalmitoylphosphatidylcholine studied in monomolecular layers

Two-component monomolecular layers were formed with DPPC and two stereoisomers of zeaxanthin 9- cis and 13- cis at the argon–water interface. Very distinct over-additivity which represents affection of a lipid arrangement in the membrane has been observed in the case of zeaxanthin 9- cis (maximum at 20 mol%) but not in the case of zeaxanthin 13- cis. The differences in the organization of the isomers of zeaxanthin–DPPC monolayers are interpreted in terms of the different orientation of both xanthophylls at the interface observed at relatively high surface pressures (>25 mN/m) comparable to the surface pressures of biomembranes. The results are consistent with the model according to which zeaxanthin 9- cis adopts a vertical orientation at the polar–nonpolar interface in contrast to zeaxanthin 13- cis, which is oriented horizontally owing to the fact that it interacts by two hydroxyl groups with the same hydrophobic–hydrophilic interface in the monolayer. The findings are discussed in comparison with the behavior of zeaxanthin in the conformation all- trans in the same system. Zeaxanthin all- trans forms efficiently molecular aggregates in the mixed monolayers in contrast to cis isomers. Circular dichroism measurements show the formation of molecular structures by zeaxanthin 13- cis that are interpreted as dimers. FTIR measurements show that these dimers are stabilized by van der Waals interactions unlike aggregated structures formed by all- trans zeaxanthin that are stabilized by hydrogen bonding. Physiological importance of the differences in aggregation and orientation of stereoisomers of zeaxanthin in lipid environment is discussed.

Interaction between chlorophyll a and violaxanthin in different steric conformations : Model studies in monomolecular layers

Monomolecular layers of pure chlorophyll a (Chl a), violaxanthin in the conformation all- trans, 9- cis, 13- cis and two-component monomolecular layers formed with Chl a and all isomers of violaxanthin, at a stoichiometric ratio 1:1, were formed and compressed at the argon–water interface. The specific molecular areas of Chl a and the isomers of violaxanthin in monomolecular layers were determined on the basis of the isotherms of compression. The isotherms of compression of the Chl a–violaxanthin-all trans monolayers correspond well to the theoretical ones, calculated on the basis of the additivity rule. The theoretical isotherms of compression of monolayers formed with Chl a and violaxanthin in conformations 9- cis and 13- cis are shifted towards higher molecular areas with respect to the experimental ones, indicating molecular interaction between components of the system. Monolayers were also deposited to a solid support by means of the Langmuir–Blodgett technique and subjected to spectroscopic studies: to quartz lamella in the case of electronic absorption and fluorescence spectroscopy and to ZnSe crystal in the case of Fourier Transform Infrared Spectroscopy (FTIR). The comparison of the absorption and Chl a fluorescence excitation spectra enabled to make a calculation of the rate of excitation energy transfer from violaxanthin to Chl a. Efficiency of energy transfer from a carotenoid to Chl a was 32% in the case of violaxanthin all- trans, 21% in the case of violaxanthin 9- cis and 15% for violaxanthin 13- cis. Such findings suggest a structural role of violaxanthin in conformation 9- cis and 13- cis and an accessory role of violaxanthin all- trans in the photosynthetic apparatus. The analysis of FTIR spectra reveals molecular interaction between Chl a and violaxanthin in conformations cis via the hydroxyl groups of xanthophylls located in the C3 and C3′ positions.

Polyene antibiotic amphotericin B in monomolecular layers: spectrophotometric and scanning force microscopic analysis

Monolayers of amphotericin B (AmB) and monolayers composed of AmB and dipalmitoylphosphatidylcholine (DPPC) were formed at the argon–water interface and deposited on a solid support by means of the Langmuir–Blodgett technique. The hypsochromic shift observed in the absorption spectra of monolayers is indicative of aggregated structures of AmB. The exciton splitting theory allowed us to calculate the distance between neighboring molecules in the aggregates as 7.8 Å. Scanning force microscopy of the AmB monolayers revealed the formation of a homogeneous monolayer composed of molecules separated by a distance of 6–8 Å. Microscopy also reveals the formation of cylindrical structures of AmB with a diameter close to 17 Å (internal diameter close to 6 Å) in the monolayers containing additionally 10 mol% DPPC.

Organization of mixed monomolecular layers formed with the xanthophyll pigments lutein or zeaxanthin and dipalmitoylphosphatidylcholine at the argon–water interface

Two-component monomolecular layers were formed with two xanthophyll pigments, lutein and zeaxanthin and dipalmitoylphosphatidylcholine (DPPC), at the argon–water interface. Analysis of the mean molecular area parameters versus molar fraction of the xanthophyll component shows large overadditivity (ca. 50 Å 2 in the case of zeaxanthin and 150 Å 2 in the case of lutein) in the region of low molar fractions of carotenoids (maximum at 5 mol% in the case of zeaxanthin and at 20 mol% in the case of lutein). The experimental values of a mean molecular area are in good agreement with the values expected, based on the additivity rule at high molar percentages of the xanthophylls. Absorption spectroscopy of a single monolayer at the argon–water interface in the UV–Vis region has also been applied to analyze the formation of molecular assemblies of lutein in monomolecular films. The differences in the organization of lutein–DPPC and zeaxanthin–DPPC monolayers are interpreted in terms of the aggregation of xanthophyll pigments in the layers and different orientation of both xanthophylls at the interface. The results are discussed in relation to possible physiological functions of lutein and zeaxanthin in the membranes of the retina of an eye.

Xanthophyll pigments in light-harvesting complex II in monomolecular layers: localisation, energy transfer and orientation

Monomolecular layers of the largest light-harvesting pigment-protein complex of Photosystem II (LHCII) were formed at the argon-water interface. The molecular area of the LHCII monomer in monomolecular layers determined from the isotherms of compression is found to be close to 14 nm 2, which corresponds well to the molecular dimensions of the protein evaluated on the basis of crystallographic studies. Monolayers of LHCII were deposited on a glass support by means of the Langmuir-Blodgett technique and subjected to spectroscopic studies: electronic absorption spectrophotometry and spectrofluorometry. The fluorescence excitation spectra of chlorophyll a in monolayers of LHCII were analysed using gaussian deconvolution. Comparison of the absorption and fluorescence excitation spectra enabled calculation of the rate of excitation energy transfer in the system. Excitation energy was found to be transferred to chlorophyll a from chlorophyll b with 97% efficiency, from neoxanthin with 85%, from lutein with 62% and from violaxanthin with at least 54% efficiency. The analysis of the position of the 0-0 absorption band of the xanthophylls revealed that neoxanthin is located in the same protein environment as lutein but in a different environment than violaxanthin. The analysis of fluorescence excitation spectra of chlorophyll a in LHCII, recorded with the excitation light beam polarised in two orthogonal directions, enabled the determination of the mean orientation angle of the accessory xanthophyll pigments with respect to the plane of the sample. The mean orientation of lutein found in this study (approx. 51°) corresponds well to the crystallographic data. Neoxanthin was found to adopt a similar orientation to lutein. The transition dipole moment of violaxanthin was found to form a mean angle of 71° with the axis spanning two polar regions of the protein, perpendicular to the plane of the monolayer, suggesting planar orientation of this pigment with respect to the plane of the thylakoid membrane. These experimentally determined xanthophyll orientations are discussed in terms of importance of peripheral xanthophyll pigments in supramolecular organisation of LHCII and the operation of the xanthophyll cycle within the thylakoid membrane.

Light-induced excitation quenching and structural transition in light-harvesting complex II

Light-induced fluorescence quenching of chlorophyll a in light-harvesting complex II (LHCII) incorporated into liposomes was examined. The rate of fluorescence quenching was found to depend on the incubation temperature. The effect was almost not observed at liquid nitrogen temperature, demonstrated a lag phase after onset of light at temperatures below 25 °C and was most distinctly pronounced at temperatures above 25 °C. Energetic uncoupling of accessory xanthophylls and chlorophyll a, and energetic uncoupling of chlorophyll b and chlorophyll a were observed as accompanying the excitation quenching. The observed changes were reversible during dark incubation. Similar energetic uncoupling was also observed in darkness, induced by the increase in temperature. Additionally, the temperature characteristics of fluorescence measurements displayed a pronounced transition in the region of 22–25 °C. The experiments carried out with the monomolecular layer technique in dicated a structural transition of LHC II in the same temperature region as demonstrated by an increase in the mean molecular area of LHC II at the argon-water interface. Alterations in surface topography induced by temperature changes could also be observed with scanning force microscopy of LHC II monolayers deposited as Langmuir-Blodgett films onto glass slides. The transition was found to be associated with the enhanced excitation energy consumption by the protein, monitored by calorimetric measurements. It is proposed that the observed transition efficiently protects LHC II against overexcitation-related damage and is therefore of physiological importance.

Surface behaviour of long-chain lipolytic products (a 1-to-1 mixture of oleic acid and diolein) spread as monomolecular films in the presence of long-chain triglycerides

Surface pressure-molecular area curves of pure and mixed monolayers of lipolytic products (a mixture of oleic acid and 1,2-dioleoyl- sn-glycerol (OA-DO) at a 1:1 molar ratio), oleic acid (OA) and purified soybean oil (TG) spread at the argon-water interface were obtained. The results reveal that OA-TG and (OA-DO)-TG mixtures behave ideally when spread over a subphase at pH 8.0. We also determined the surface pressure-molecular area curves of lipolytic products (OA-DO) at the argon-water interface in the absence or presence of an excess of triglycerides. In the presence of an excess of TG, the films of OA-DO can be compressed up to surface pressures identical to the collapse pressure of the OA-DO when present alone. Furthermore, the fact that at the collapse pressure, the area occupied by the OA-DO mixture was the same in both the absence and presence of TG showed that under the experimental conditions used, negligible amounts of lipolytic products were dissolved in the oil and aqueous phases at both pH 5 and pH 8. The fraction of the surface occupied by OA-DO molecules varied from 80 to 100% at surface pressures ranging from 18 to 30 mN m −1.

Regulation of fatty acid 18O exchange catalyzed by pancreatic carboxylester lipase. 2. Effects of lateral lipid distribution in mixtures with phosphatidylcholine.

The lipase-catalyzed exchange of the carboxyl oxygens of 13,16-cis,cis-docosadienoic acid (DA) was studied in the presence of a nonsubstrate matrix lipid, 1-palmitoyl-2-oleoylphosphatidylcholine. For mixed lipid films at the argon-water interface exposed to pancreatic carboxylester lipase (EC 1.1.1.13), the extent of oxygen exchange showed an abrupt increase as the abundance of DA in the interface was increased from 0.5 to 0.6 mole fraction. This compositional range was independent of the level of enzyme used and of the surface pressure, i.e., lipid packing density, of the film. Concomitant with the transition was a change in the apparent mechanism of exchange from coupled to random sequential. Like the extent of oxygen exchanged, the shift in mechanism was independent of all variables except the lipid composition of the interface. The absence of any chemical or physical change accompanying the exchange reaction precludes mechanistic explanations based on the generation of reaction products by the enzyme. Instead, the results suggest that the lateral distribution of DA in phosphatidylcholine-DA interfaces regulates the expression of carboxylester lipase activity and its apparent mechanism. Preliminary measurements give an average cluster size of 1825 molecules of DA when its mole fraction is 0.35. As the DA content of the interface reaches 0.5-0.6, there appears to be a lipid head-group based percolative transition in which DA becomes the continuum. Because this transition involves the lateral organization of the lipids themselves, other interfacially active enzymes may be regulated similarly.

Surface dipole moments of lipids at the argon-water interface. Similarities among glycerol-ester-based lipids

Surface potential-surface pressure-area isotherms at the argon-buffer interface have been determined for 38 lipid species comprising 19 chemical classes. These lipids all exhibited a finite range of liquid-expanded surface pressure-area behavior. For most species, the linearity of surface potential with reciprocal area was excellent, but nonzero intercepts were obtained. This suggests a lipid-induced reorganization of interfacial water molecules which is area independent. The linearity of the data permits calculation of the surface dipole moment, mu perpendicular, for each lipid. The values of mu perpendicular for a series of oleoyl-containing acylglycerols, dioleoyl phosphatidylcholine, and dioleoyl phosphatidylethanolamine exhibit acylglycerol ester group mu perpendicular's which are generally consistent with known conformational properties of such lipids. The values are 132 mD for the perpendicular oleoyl glycerol-ester group and 252 mD for that in the kinked-chain conformation. Comparison of mu perpendicular's calculated using these values with homologues confirms the approximate independence of mu perpendicular from aliphatic chain length and permits identification of exceptions with possible conformational or orientational differences. Notably, diphytanoyl phosphatidylcholine shows a 45% larger mu perpendicular than predicted. Differences in mu perpendicular among lipid classes allow estimation of the electrical consequences of lipid metabolism and exchange. Calculations show that reactions such as the generation of 1,2-diacylglycerol from diacyl glycerophosphocholine or diacyl glycerophosphoinositol should produce surface potential changes of -127 and +42 mV, respectively. Thus, the two phospholipids are not simply alternative sources of diacylglycerol with respect to processes dependent on surface potential.

Miscibility, chain packing, and hydration of 1-palmitoyl-2-oleoyl phosphatidylcholine and other lipids in surface phases

The miscibility of 1-palmitoyl-2-oleoyl phosphatidylcholine with triolein, 1,2-diolein, 1,3-diolein, 1(3)-monoolein, oleyl alcohol, methyl oleate, oleic acid, and oleyl cyanide (18:1 lipids) was studied at the argon-water interface. The isothermal phase diagrams for the mixtures at 24 degrees were characterized by two compositional regions. At the limit of miscibility with lower mol fractions of 18:1 lipid, the surface pressure was composition-independent, but above a mixture-specific stoichiometry, surface pressure at the limit of miscibility was composition-dependent. From the two-dimensional phase rule, it was determined that at low mol fractions of 18:1 lipids, the surface consisted of phospholipid and a preferred packing array or complex of phospholipid and 18:1 lipid, whereas, above the stoichiometry of the complex, the surface phase consisted of complex and excess 18:1 lipids. In both regions of the phase diagram, mixing along the phase boundary was apparently ideal allowing application of an equation of state described earlier (J. M. Smaby and H. L. Brockman, 1984, Biochemistry, 23:3312-3316). From such analysis, apparent partial molecular areas and hydrations for phospholipid, complex, and 18:1 lipid were obtained. Comparison of these calculated parameters for the complexed and uncomplexed states shows that the aliphatic moieties behave independently of polar head group. The transition of each 18:1 chain to the complexed state involves the loss of about one interfacial water molecule and its corresponding area. For 18:1 lipids with more than one chain another two water molecules per additional chain are present in both states but contribute little to molecular area. In contrast to 18:1 lipids, the phospholipid area and hydration change little upon complexation. The uniformity of chain packing and hydration behavior among 18:1 lipid species contrasts with complex stoichiometries that vary from 0.04 to 0.65. This suggests that the stoichiometry of the preferred packing array is determined by interactions involving the more polar moieties of the 18:1 lipids and the phospholipid.

Lipolysis and lipid movement in a membrane model. Action of lipoprotein lipase.

The action of purified bovine milk lipoprotein lipase on tri[3H]oleoylglycerol and the effect of albumin on movement of lipolytic products at an argon-water interface were studied in a specially designed tricomparted trough. The amount of trioleoylglycerol applied was 14 times that needed to cover the surface of the aqueous subphase (0.1 M Tris . HCl, pH 7.4) with a monolayer. It is concluded that trioleoylglycerol was present in lenses on the surface of the aqueous subphase, that hydrolysis by lipoprotein lipase occurred in or near the lipid/argon-water interface, and that lipolytic products immediately located and spread throughout the interface, displacing substances with lower spreading pressures from the interface. Addition of albumin to the aqueous subphase accelerated markedly the desorption of oleic acid and monooleoylglycerol from the interface and thereby enhanced lipolysis. When albumin was not contiguous with the site of hydrolysis, oleic acid and monooleoylglycerol readily moved in the interface to the area of contact with albumin where they were desorbed from the interface. These findings support the hypothesis of transport of lipolytic products by lateral movement in cell membranes.