Cytochrome c induced lateral phase separation in a diphosphatidylglycerol-steroid spin-label model membrane

Birrell, G. Bruce; Griffith, O. Hayes

doi:10.1021/bi00658a035

Cited by 112 publications

(33 citation statements)

References 23 publications

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“…In a related 31P N M R study of cytochrome c complexes with twocomponent lipid bilayers containing cardiolipin and DOPC (Spooner & Watts, 1992), no enhancement of the 31P spinlattice relaxation for the cardiolipin component was reported, in contrast with the results observed for the complexes of cytochrome c with single-component lipid bilayers (Spooner & Watts, 1991;Pinheiro & Watts, 1994). However, some perturbation of the cardiolipin component by the protein in mixed DOPC/CL bilayers may occur, in agreement with suggestions that possible phase separation of cardiolipin in mixed lipid membranes is induced upon binding of cytochrome c, as shown by spin-label ESR (Birrel & Griffith, 1976) and * H N M R studies (Brown & Wuthrich, 1977). Based solely on the relative intensities of the isotropic resonances in the central band of 31P MAS N M R spectra of DOPC/CL bilayers with and without cytochrome c, a reduction in the chemical shift anisotropy of the cardiolipin component in this binary bilayer was predicted, indicating some selective influence of cytochrome c on CL.…”

Section: Recalling Basic Nmr Concepts the Dipolar Interaction (D)supporting

confidence: 88%

Resolution of Individual Lipids in Mixed Phospholipid Membranes and Specific Lipid-Cytochrome c Interactions by Magic-Angle Spinning Solid-State Phosphorus-31 NMR

Pinheiro¹,

Watts²

1994

Biochemistry

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A model of the inner mitochondrial membrane was constructed with dioleoyphosphatidylcholine (PC), dioleoylphosphatidylethanolamine (PE), and cardiolipin (CL) at a PC:PE:CL molar ratio of 2:2:1, and the interaction of the peripheral membrane protein cytochrome c with this mixed membrane has been investigated by static and magic-angle spinning (MAS) solid-state 31P NMR. The static 31P NMR spectrum of the three-component membrane is a typical broad powder pattern for phospholipids in a bilayer structure, and is a result of three overlapping spectra of each individual phospholipid component in the mixed membrane, with an average effective chemical shift anisotropy of approximately 41 ppm. Using magic-angle spinning NMR methods, three resolved resonances are observed in the narrowed MAS 31P NMR spectrum, each of which has been assigned to each lipid component in the mixed membrane. This allows the investigation of individual phospholipid-protein interactions in multicomponent lipid bilayers. The interaction of cytochrome c with each lipid in a model mitochondrial membrane could now be evaluated. Phosphorus-31 spin-lattice (T1) relaxation times for each lipid phosphate were measured as a function of temperature, in the absence and presence of bound cytochrome c. T1 was not affected for any lipid upon binding of cytochrome c over the temperature range analyzed. However, averaging of the phosphorus-31 chemical shift anisotropy for the cardiolipin component in mixed PC/PE/CL bilayers at lower temperatures ceases to be axially symmetric on binding of cytochrome c, while for PC and PE components the axial symmetry is retained over the temperature interval studied here.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Recalling Basic Nmr Concepts the Dipolar Interaction (D)supporting

confidence: 88%

Resolution of Individual Lipids in Mixed Phospholipid Membranes and Specific Lipid-Cytochrome c Interactions by Magic-Angle Spinning Solid-State Phosphorus-31 NMR

Pinheiro¹,

Watts²

1994

Biochemistry

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“…The ability of cytochrome c to bind to acidic phospholipids and to induce phase separations in acidic phospholipid containing vesicles is well documented (Gorrissen et al, 1986;de Kruijff and Cullis, 1980;Birrell and Griffith, 1976;Killian and de Kruijff, 1986;Mannella et al, 1987;Mustonen et al, 1987;Rietveld et al, 1986) although not all studies have demonstrated that phase separations are induced . In the present report, advantage was taken of methods for the visualization of domain formation using fluorescence microscopy to demonstrate that not only did cytochrome c bind to acidic phospholipids, but such interactions led to the formation of a large PA domain within the vesicle bilayer (Fig.…”

Section: Discussionmentioning

confidence: 99%

Influence of proteins on the reorganization of phospholipid bilayers into large domains

Haverstick

Gläser

1989

Biophysical Journal

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Using large (5-10 microns) vesicles formed in the presence of phospholipids fluorescently labeled on the acyl chain and visualized using a fluorescence microscope, charge-coupled-device camera, and digital image processor, we examined the effects of membrane proteins on phospholipid domain formation. In vesicles composed of phosphatidic acid and phosphatidylcholine, incubation with cytochrome c induced the reorganization of phospholipids into large phosphatidic acid-enriched domains with the exclusion of phosphatidylcholine. Cytochrome c binding was demonstrated to be highest in the phosphatidic acid-enriched domain of the vesicle using the absorbance of the heme moiety for visualization. Both binding of cytochrome c and phospholipid reorganization were blocked by pretreatment of the vesicles with 0.1 M NaCl. The pore forming peptide gramicidin was examined for the effects of an integral protein on domain formation. Initially, gramicidin distributed randomly within the vesicle and showed no phospholipid specificity. Phosphatidic acid domain formation in the presence of 2.0 mM CaCl2 or 100 microM cytochrome c was not affected by the presence of 5 mol % gramicidin within the vesicles. In both cases, gramicidin was preferentially excluded from the phosphatidic acid-enriched domain and became associated with phosphatidylcholine-enriched areas of the vesicle. Thus, cytochrome c caused a major reorganization of both the phospholipids and the proteins in the bilayer.

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“…A similar phenomenon occurs when cytochrome c, or other basic proteins, bind to membranes containing acidic phospholipids (15)(16)(17)(18)(19)(20)(21)(22).…”

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confidence: 90%

Characterization of lipid domains in erythrocyte membranes.

Rodgers

Gläser

1991

Proc. Natl. Acad. Sci. U.S.A.

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Fluorescence digital imaging microscopy was used to study the lateral distribution of the lipid components in erythrocyte membranes. Intact erythrocytes labeled with phospholipids containing a fluorophore attached to one fatty acid chain showed an uneven distribution ofthe phospholipids in the membrane thereby demonstrating the presence of membrane domains. The enrichment of the lipotropic compound chlorpromazine in domains in intact erythrocytes also suggested that the domains are lipid-enriched regions. Similar membrane domains were present in erythrocyte ghosts. The phospholipid enrichment was increased in the domains by inducing membrane protein aggregation. Double-labeling experiments were done to determine the relative distributions of different phospholipids in the membrane. Vesicles made from extracted lipids did not show the presence of domains consistent with the conclusion that membrane proteins were responsible for creating the domains. Overall, it was found that large domains exist in the red blood cell membrane with unequal enrichment of the different phospholipid species.Since it was proposed almost 20 years ago, the Fluid Mosaic Model has served as a foundation for understanding membrane structure and function (1). The model describes biological membranes as a mixture of lipid and protein components in a phospholipid bilayer where all the components can undergo free lateral motion. This results in a random distribution of the membrane components with no long-range order. One refinement to this model is the role of the cytoskeleton in anchoring transmembrane proteins (2). Another change is that biological membranes may be more heterogeneous than depicted by the Fluid Mosaic Model with the existence of relatively large domains with compositions different from the bulk membrane.

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Cytochrome c induced lateral phase separation in a diphosphatidylglycerol-steroid spin-label model membrane

Cited by 112 publications

References 23 publications

Resolution of Individual Lipids in Mixed Phospholipid Membranes and Specific Lipid-Cytochrome c Interactions by Magic-Angle Spinning Solid-State Phosphorus-31 NMR

Resolution of Individual Lipids in Mixed Phospholipid Membranes and Specific Lipid-Cytochrome c Interactions by Magic-Angle Spinning Solid-State Phosphorus-31 NMR

Influence of proteins on the reorganization of phospholipid bilayers into large domains

Characterization of lipid domains in erythrocyte membranes.

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