Fourier transform infrared spectroscopic identification of gel phase domains in reconstituted phosphilipid vesicles containing Ca2+-ATPase

Jaworsky, Markian S; Mendelsohn, Richard

doi:10.1016/0005-2736(86)90546-8

Cited by 12 publications

(7 citation statements)

References 33 publications

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“…Since the change in the DEPC band shape in the 950-1000 cm™1 spectral region at the 0.7-kbar transition is identical with that observed at the temperature-induced transition (Jaworsky & Mendelsohn, 1986), we anticipate that the temperatureinduced transition in DOPC will also be accompanied by significant changes in the band shape of the out-of-plane CH wagging mode.…”

Section: Resultsmentioning

confidence: 61%

Effects of cis and trans unsaturation on the structure of phospholipid bilayers: a high-pressure infrared spectroscopic study

Siminovitch

Wong²,

Mantsch³

1987

Biochemistry

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In order to compare the effects of cis and trans unsaturation on the structure and packing of phospholipid bilayers, infrared spectra of aqueous dispersions of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dielaidoyl-sn-glycero-3-phosphocholine (DEPC) were measured in a diamond anvil cell at 28 degrees C as a function of pressure up to 36 kbar. The infrared spectra indicate that DEPC and DOPC undergo pressure-induced liquid-crystalline to gel phase transitions at critical pressures of 0.7 and 5.2 kbar, respectively. Below their respective critical pressures, the infrared spectra of DOPC and DEPC are essentially indistinguishable, whereas above these pressures, there are very pronounced differences in the barotropic behavior of these two lipids. Specifically, at the 5.2-kbar transition in DOPC, there are significant changes in the frequencies, intensities, and widths of bands associated with the interfacial C = O groups, the olefinic CH = CH groups, and the terminal CH3 groups, whereas the corresponding bands of DEPC are, by contrast, relatively insensitive to the pressure-induced phase transition. The unusual band shape changes in DOPC are attributed to a unique packing arrangement of the oleoyl acyl chains required to accommodate the bent geometries of adjacent cis double bonds. Moreover, above 5 kbar in DEPC, well-defined correlation field splittings of the CH2 scissoring and rocking modes are observed, with magnitudes very similar to those observed at comparable pressures in saturated lipid systems. The absence of correlation field splittings of the corresponding bands of DOPC up to 36 kbar suggests that the bent oleoyl acyl chains are closely packed with all chains oriented parallel to each other.

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Section: Resultsmentioning

confidence: 61%

Effects of cis and trans unsaturation on the structure of phospholipid bilayers: a high-pressure infrared spectroscopic study

Siminovitch

Wong²,

Mantsch³

1987

Biochemistry

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“…description, i.e., exclusion of the higher melting compoe that the positions of maximum intensity nent, has been invoked for phase-separated lipid systems shifts from the true peak values because of reconstituted with CaATPase (12). However, for the folterference from liquid water absorption near lowing reasons, the explanation is incomplete in the current rhe pair of features at 1,625 and 1,680 cm-' system: (a) When CaATPase interacts with pure DEPC, to f3-antiparallel sheet secondary structure the melting of the lipid is considerably reduced in temperafor assignments), whereas the band at 1,660 ture from the pure lipid (13). In the current case, the from a-helical amide I modes.…”

Section: Discussion Igh Temperature Transitions and A Resultantmentioning

confidence: 79%

“…Four lipid-protein complexes of different overall lipid compositions were prepared and examined. The activities of the complexes are lower than have been achieved in other systems (12)(13)(14) using the same reconstitution protocols and enzyme preparations with similar starting activities. From the data in Table I, there does not appear to be a strong dependence of enzyme activity on the lipid level.…”

Section: Binary Lipid Mixturesmentioning

confidence: 66%

“…Dr. Jaworsky's current address is Department of Chemistry, University of California, Riverside, CA 92521. we have, in previous studies from this laboratory (9)(10)(11)(12)(13)(14), been involved with the reconstitution of glycophorin from the human erythrocyte membrane (9,10) and CaATPase from rabbit skeletal muscle (11)(12)(13)(14) into binary lipid mixtures. The latter are the simplest preparations that can mimic some of the in vivo possibilities and have been chosen in our studies to span a range of lipid classes and phase behaviors.…”

Section: Mendelsohnmentioning

confidence: 99%

“…(b) Selection of the lower melting component in lipid mixtures that showed partial or complete gel state phase separation (SOPC/ DPPC-d62 [14] or DOPC/DPPC-d62 [12]). (c) The ability to induce gel state phase separation in a complex lipid 241 environment (i.e., DEPC/DMPC-dM/native lipids mixture) (13).…”

Section: Mendelsohnmentioning

confidence: 99%

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Unusual partitioning behavior of CaATPase in dipalmitoylphosphatidylethanolamine/dielaidoylphosphatidylcholine++ + mixtures

Jaworsky

Mendelsohn

1987

Biophysical Journal

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CaATPase from rabbit sarcoplasmic reticulum has been isolated, purified, stripped of its native lipids, and reconstituted into binary lipid mixtures of dielaidoylphosphatidylcholine (DEPC) and dipalmitoylphosphatidylethanolamine (DPPE) or acyl-chain perdeuterated DPPE (DPPE-d62). The partitioning properties of the protein were determined from differential scanning calorimetry (DSC) and Fourier transform infrared (FT-IR) spectroscopy. Acyl-chain perdeuteration allows the separate determination of the order and melting characteristics of each lipid species with FT-IR. The binary lipid mixture has been shown to be phase separated in the gel state (Brauner, J. W., and R. Mendelsohn, 1986, Biochim. Biophys. Acta, 861:16-24). The solid phases present at low temperatures correspond to a pure DEPC phase and a mixed phase of DEPC/DPPE-d62. Insertion of protein at 37 degrees C leads to a domain of relatively protein-free DPPE-d62 and a phase containing both lipids plus CaATPase. We suggest that CaATPase selects a fixed composition (60% DEPC, 40% DPPE-d62) for its immediate environment. The composition of the lipid in the immediate vicinity of protein is largely independent of the initial DEPC/DPPE-d62 ratios in the reconstitution protocol. The relevance of these results to observations of discrete domains in native membranes is discussed.

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A SANS Study of High Pressure Phase Transitions in Model Biomembranes

Winter

Pilgrim

1989

Ber Bunsenges Phys Chem

142

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The phase behaviour and thermotropic and barotropic phase transitions of different model biomembranes (DPPC, DMPC, DEPC and POPC*)), which consist of different hydrocarbon chain length and degree of unsaturation, have been investigated by small‐angle neutron scattering (SANS) as a function of temperature and pressure up to about 65°C and 3 kbar, respectively. A pressure induced structural phase transition from a liquid‐crystalline to a gel state is observed for the whole temperature range studied. The pressure of this transition increases with increasing temperature with a slope dTm/dP of about 21°C/kbar, which is common for all the different lipids investigated. The SANS experiments indicate that the effect of the two trans double bonds in DEPC and the cis double bond in the sn‐2 hydrocarbon chain of POPC enhances the conformational disorder in the hydrocarbon chains not only in the liquid‐crystalline, but also in the gel state. For DEPC and POPC the prefered structural conformation in the gel state is probably not tilted as it is in the gel state of the saturated phospholipids DMPC and DPPC. Only for DPPC multilamellar vesicles, an additional gel phase at higher pressures has been observed, which can be ascribed to an interdigitation of the hydrocarbon chains of opposing chains in the lipid bilayer system.

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Fourier transform infrared spectroscopic identification of gel phase domains in reconstituted phosphilipid vesicles containing Ca2+-ATPase

Cited by 12 publications

References 33 publications

Effects of cis and trans unsaturation on the structure of phospholipid bilayers: a high-pressure infrared spectroscopic study

Effects of cis and trans unsaturation on the structure of phospholipid bilayers: a high-pressure infrared spectroscopic study

Unusual partitioning behavior of CaATPase in dipalmitoylphosphatidylethanolamine/dielaidoylphosphatidylcholine++ + mixtures

A SANS Study of High Pressure Phase Transitions in Model Biomembranes

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