Spin-label electron spin resonance study of bacteriophage M13 coat protein incorporation into mixed lipid bilayers

Datema, K.P.; Wolfs, C.J.A.M.; Marsh, Derek; Watts, Anthony; Hemminga, M.A.

doi:10.1021/bi00398a006

Cited by 32 publications

(32 citation statements)

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“…In the absence of lipid the 50% saturation was achieved after 2.4 x 105 sec in the absence of lipids and after 8.7 x 104 in the presence of the lipid (Fig. 4 A and C) (15,41) or activation of lipases (36). One can speculate that class II MHC molecules may be found in different conformers within the cell, attaining an active binding state under the influence of component specific to subcellular microenvironments-e.g., in endosomes.…”

Section: Resultsmentioning

confidence: 91%

Phospholipids enhance the binding of peptides to class II major histocompatibility molecules.

Roof

Luescher²,

Unanue³

1990

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

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The binding of a lysozyme and ovalbumin peptide to purified class II major histocompatibiity molecules in detergents was increased by the addition of certain lipids. Natural lipids from B lymphoma cells enhanced the binding and so did phosphatidylcholine, phosphatidylserine, phosphatidylinositol, and cardiolipin. Phosphatidylethanolamine, sphingomyelin, and cholesterol had no effect. There was no major difference between the effects of a phospholipid and its lyso derivative. As studied with phosphatidylcholine, the increase in peptide binding was also dependent on the fatty acid composition of the lipid. (MHC) proteins (1,2). The extent of binding of peptides to histocompatibility molecules, which can be studied in detergent solutions (1-7), usually correlates with their immunogenicity in vivo (2-4). There are, however, indications that the interactions of peptides with class II MHC may be quantitatively different on live antigen-presenting cells. For example, the binding of antigenic peptides to class II MHC molecules on live cells was reported to be much faster than that found in solution (8-10). A similar result was found for the dissociation of peptides bound to the class II MHC molecules (11).We recently found that the labeling of class II MHC molecules to peptides containing a photoreactive compound was influenced by phospholipids, both in the extent as well as in the relative degree ofthe a or p chain labeling (12). When tested on cell membranes that contained class II MHC proteins, photoreactive conjugates of the hen egg white lysozyme (HEL) peptide from residues 46 to 61 labeled exclusively the a chain of I-Ak (13). Conversely, I-Ak purified in an equimolar solution of MEGA 8 and MEGA 9 detergents was labeled to similar extent on both chains. However, the addition of certain lysophospholipids resulted in strong binding, with lysophosphatidylcholine predominantly labeling the a chain and lysophosphatidylserine labeling the ,B chain. We concluded that class II MHC molecules contained dialyzable components required for their binding integrity and that, most likely, these components were lipids.Lipid-protein interactions are of critical importance in maintaining the biological activity of a number of membrane proteins (for review, see refs. 14-16). These interactions may have different degrees of specificity and often induce conformational changes in the protein critical for their function (14,(17)(18)(19)(20)(21)(22). Lipids, by interacting directly with the MHC protein or by changing the structure ofthe detergent micelles, may induce conformational changes reflected in the distal binding site of the protein (23,24). Consistent with this hypothesis are the observations indicating that class II MHC molecules reconstituted in lipid monolayers present peptides differently, depending on the lipid composition (25). Other studies have shown that treatment of antigen-presenting cells with purified phospholipase A2 or C, or with cerulenin, an antibiotic that interferes with lipid metabolism, greatly dimi...

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Section: Resultsmentioning

confidence: 91%

Phospholipids enhance the binding of peptides to class II major histocompatibility molecules.

Roof

Luescher²,

Unanue³

1990

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

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“…A Digital Equipment Corp. LPS system and a dedicated PDP 11/10 computer with a VTll display were used to digitize the spectra. ESR spectrometer settings were those given in Datema et al (1987a).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, we have been able to incorporate M13 coat protein also in pure DMPC lipids so that now the lipid-protein interaction can be studied as a function of increasing content of negatively charged phospholipid. This is especially interesting, since in earlier experiments (Datema et al, 1987a) the protein was shown to have an increased specificity for negatively charged phospholipids.The present paper examines the effect of M 13 coat protein on mixed systems with variable amounts of DMPG and DMPC. The number of lipid association sites is determined in pure DMPC, as well as the relative association constants, for one representative spin-labeled phospholipid from each of the above three groups of phospholipids of different specificity.…”

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confidence: 87%

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confidence: 87%

“…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).…”

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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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Application of Fluorescence to Understand the Interaction of Peptides with Binary Lipid Membranes

Almeida¹,

Loura

Prieto³

2005

Reviews in Fluorescence 2005

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Spin-label electron spin resonance study of bacteriophage M13 coat protein incorporation into mixed lipid bilayers

Cited by 32 publications

References 20 publications

Phospholipids enhance the binding of peptides to class II major histocompatibility molecules.

Phospholipids enhance the binding of peptides to class II major histocompatibility molecules.

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

Application of Fluorescence to Understand the Interaction of Peptides with Binary Lipid Membranes

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