Spin-label studies of lipid-protein interactions in sodium-potassium ATPase membranes from rectal glands of Squalus acanthias

Esmann, Mikael; Watts, Anthony; Marsh, Derek

doi:10.1021/bi00327a016

Cited by 78 publications

(50 citation statements)

References 38 publications

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“…Although one of the major thylakoid lipid components, namely, monogalactosyldiacylglycerol, is incapable alone of forming lamellar membrane structures, the basic features of the lipid-protein interactions remain the same in the lamellar membranes of intact thylakoids and subthylakoid membrane fragments as in other biomembranes. Similar results have been found in other membranes that contain nonlamellar forming phospholipids, for example, the rod outer segment disk mem- brane (Watts et al, 1979;Mollevanger & De Grip, 1984), Na,K-ATPase membranes (Esmann et al, 1985;D. Marsh and M. Esmann, unpublished data), and cytochrome oxidase reconstituted with cardiolipin (Powell et al, 1985, and unpublished data).…”

Section: Discussionsupporting

confidence: 78%

Spin-label ESR studies of lipid-protein interactions in thylakoid membranes

Li¹,

Knowles²,

Murphy³

et al. 1989

Biochemistry

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Lipid-protein interactions in thylakoid membranes, and in the subthylakoid membrane fractions containing either photosystem 1 or photosystem 2, have been studied by using spin-labeled analogues of the thylakoid membrane lipid components, monogalactosyldiacylglycerol, phosphatidylglycerol, and phosphatidylcholine. The electron spin resonance spectra of the spin-labeled lipids all consist of two components, one corresponding to the fluid lipid environment in the membranes and the other to the motionally restricted membrane lipids interacting directly with the integral membrane proteins. Spectral subtraction has been used to quantitate the fraction of the membrane lipids in contact with the membrane proteins and to determine the selectivity between the different lipid classes for the lipid-protein interaction. The fractions of motionally restricted lipid in the thylakoid membrane are 0.36, 0.39, and 0.53, for the spin-labeled monogalactosyldiacylglycerol, phosphatidylcholine, and phosphatidylglycerol, respectively. Spin-labeled monogalactosyldiacylglycerol exhibits very little preferential interaction over phosphatidylcholine, which suggests that part of the role of monogalactosyldiacylglycerol in thylakoid membranes is structural, as is the case for phosphatidylcholine in mammalian membranes. Spin-labeled phosphatidylglycerol shows a preferential interaction over the corresponding monogalactosyldiacylglycerol and phosphatidylcholine analogues, in contrast to the common behavior of this lipid in mammalian systems. This pattern of lipid selectivity is preserved in both the photosystem 1 and photosystem 2 enriched subthylakoid membrane fractions.T e photosynthetic apparatus of green plants comprises a series of integral protein complexes embedded in the thylakoid membrane. The conversion of light energy into a chemically useful form takes place at two reaction centers, photosystem 1 (PSI)' and photosystem 2 (PS2), which consist of complexes of different integral proteins. In the thylakoids from the mesophyll cells of higher plants, the two reaction centers and associated protein complexes are separately located in the appressed (PS2) and nonappressed (PSI) membrane regions. The appressed membranes are arranged in stacks and are interconnected by the nonappressed membranes, which contain regions of high membrane curvature. Within the thylakoid,

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Section: Discussionsupporting

confidence: 78%

Spin-label ESR studies of lipid-protein interactions in thylakoid membranes

Li¹,

Knowles²,

Murphy³

et al. 1989

Biochemistry

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“…It is known from studies of lipid-protein interactions with conventional spin-label EPR (27,33) that fully ionized fatty acids display a high affinity for interaction with the Na,K-ATPase, as compared with a background membrane lipid such as phosphatidylcholine. Consequently, ESEEM intensities of negatively charged spin-labeled fatty acids in native Na,K-ATPase membranes contain a strong contribution from D 2 O in the vicinity of the protein interface, in addition to the background contribution from the spin-labeled fatty acid in the lipid bilayer regions of the membrane.…”

Section: Resultsmentioning

confidence: 99%

Water Penetration Profile at the Protein-Lipid Interface in Na,K-ATPase Membranes

Bartucci

Guzzi

Esmann

et al. 2014

Biophysical Journal

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The affinity of ionized fatty acids for the Na,K-ATPase is used to determine the transmembrane profile of water penetration at the protein-lipid interface. The standardized intensity of the electron spin echo envelope modulation (ESEEM) from (2)H-hyperfine interaction with D2O is determined for stearic acid, n-SASL, spin-labeled systematically at the C-n atoms throughout the chain. In both native Na,K-ATPase membranes from shark salt gland and bilayers of the extracted membrane lipids, the D2O-ESEEM intensities of fully charged n-SASL decrease progressively with position down the fatty acid chain toward the terminal methyl group. Whereas the D2O intensities decrease sharply at the n = 9 position in the lipid bilayers, a much broader transition region in the range n = 6 to 10 is found with Na,K-ATPase membranes. Correction for the bilayer population in the membranes yields the intrinsic D2O-intensity profile at the protein-lipid interface. For positions at either end of the chains, the D2O concentrations at the protein interface are greater than in the lipid bilayer, and the positional profile is much broader. This reveals the higher polarity, and consequently higher intramembrane water concentration, at the protein-lipid interface. In particular, there is a significant water concentration adjacent to the protein at the membrane midplane, unlike the situation in the bilayer regions of this cholesterol-rich membrane. Experiments with protonated fatty acid and phosphatidylcholine spin labels, both of which have a considerably lower affinity for the Na,K-ATPase, confirm these results.

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“…Both the acetylcholine receptor (Marsh & Barrantes, 1978;Ellena et al, 1983) and rhodopsin Ryba et al, 1987) have been investigated in natural membranes and reconstituted with defined phosphatidylcholines. Similarly the Na+,K+-ATPase has also been studied in natural membranes and reconstituted with phosphatidylcholine (Esmann et al, 1985;Brotherus et al, 1981). Myelin proteolipid protein has also been studied in dimyristoylphosphatidylcholine recombinants .…”

Section: Discussionmentioning

confidence: 99%

“…Quantitatively, this relation between the thermodynamics of the lipid selectivity and the exchange dynamics can be seen by combining eq 1 and 2. Assuming that the on-rates are the same for the different lipids (Le., diffusion-controlled), then the ratio of the intrinsic off-rate constants for two lipids, e.g., L, and L2, is given by (Marsh, 1985;Horvfith et al, 1988) 7b-'(L2)/7b-'(LI) = Kr(Ll)/Kr(L2) (3) For the 14-PCSL, 14-PGSL, and 14-PASL spin-labeled phospholipids, the ratios of the relative association constants are 1 .O: 1.0:2.0, compared with the ratios of the inverse rate constants of 1.0:0.9:2.2, respectively. This is in good agreement with the predictions of eq 3 and further substantiates the methods used for analyzing the selectivity and exchange dynamics exhibited by the two-component lipid spectra.…”

Section: Discussionmentioning

confidence: 99%

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Spin-label ESR of bacteriophage M13 coat protein in mixed lipid bilayers. Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers

Wolfs¹,

Horváth²,

Marsh³

et al. 1989

Biochemistry

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Bacteriophage M 13 major coat protein has been incorporated at different lipid/protein ratios in lipid bilayers consisting of various ratios of dimyristoylphosphatidylcholine (DMPC) to dimyristoylphosphatidylglycerol (DMPG). Spin-label ESR experiments were performed with phospholipids labeled at the C-14 position of the sn-2 chain. For M 13 coat protein recombinants with DMPC alone, the relative association constants were determined for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels and found to be 1 .O, 1 .O, and 2.1 relative to the background DMPC, respectively. The number of association sites for each phospholipid on the protein was found to be 4 per protein monomer. The intrinsic off-rates for lipid exchange at the intramembranous surface of the protein in DMPC alone a t 30 OC were found to be 5 X lo6, 6 X lo6, and 2 X lo6 s-' for the phosphatidylcholine, phosphatidylglycerol, and phosphatidic acid spin-labels, respectively. Adding DMPG to the DMPC lipid system increased the exchange rates of the lipids on and off the protein. By gel filtration chromatography, it is found that protein aggregation is reduced after addition of DMPG to the lipid system. This is in agreement with measurements of tryptophan fluorescence, which show a decrease in quenching efficiency after introduction of DMPG in the lipid system. The results are interpreted in terms of a model relating the ESR data to the size of the protein-lipid aggregates.%e nonenveloped filamentous bacteriophage M 13, which is closely related to bacteriophages fd and fl (Ray, 1977), enters the Escherichia coli cell, leaving its major coat protein (a product of gene 8) in the cytoplasmic membrane (Marvin & Wachtel, 1975). During DNA replication and synthesis of new phage proteins, the coat protein is stored in the membrane as an integral membrane protein. Newly synthesized coat protein, which arrives at the membrane as a preprotein and is converted to the mature coat protein by the action of a leader peptidase (Kiihn et al., 1986), is stored in the membrane and encapsulates the viral DNA molecule during the membrane-associated assembly of new virus particles. The parental coat protein, after it is incorporated in the membrane, is also used for the formation of new infectious M13 bacteriophages (Armstrong et al., 1983). The coat protein of M13, whose amino acid sequence is known (Van Wezenbeek et al., 1980;Beck et al., 1978), consists of three different regions: a basic carboxyl terminus, a central hydrophobic core of 19 amino acids, and an acidic amino terminus (Chamberlain et al., 1978; Van Wezenbeek et al., 1980). Spin-label electron spin resonance (ESR)' has been shown to be a particularly useful technique for studying the interactions of M13 coat protein in reconstituted lipid systems (Datema et al., 1987a). Three groups of phospholipids could be distinguished showing different specificities for the M 13 coat protein (group I, CL and PA; group 11, PG, PS, and SA; and group 111, PC and PE). These spin-label expe...

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Spin-label studies of lipid-protein interactions in sodium-potassium ATPase membranes from rectal glands of Squalus acanthias

Cited by 78 publications

References 38 publications

Spin-label ESR studies of lipid-protein interactions in thylakoid membranes

Spin-label ESR studies of lipid-protein interactions in thylakoid membranes

Water Penetration Profile at the Protein-Lipid Interface in Na,K-ATPase Membranes

Spin-label ESR of bacteriophage M13 coat protein in mixed lipid bilayers. Characterization of molecular selectivity of charged phospholipids for the bacteriophage M13 coat protein in lipid bilayers

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