Membrane proteins in reverse micelles: myelin basic protein in a membrane-mimetic environment.
The solubility, reactivity, and conformational dynamics of myelin basic protein (MBP) from bovine brain were studied in reverse micelles of sodium bis(2-ethylhexyl) sulfosuccinate (AOT)-isooctane and water. Such a membrane-mimetic system resembles the aqueous spaces of native myelin sheath in terms of physicochemical properties as reflected in the high affinity of MBP for interfacial bound water. This is marked by the unusual profile of the solubility curve of the protein in reverse micelles, which shows optimal solubility at a much lower molar ratio of water to surfactant ([ H2O]/[AOT] = w0) than that reported for other water-soluble proteins. The role of counterions and/or charged polar head groups in the solubilization process is revealed by comparison of the solubility of MBP in nonionic surfactant micellar solutions. Whereas MBP is unfolded in aqueous solutions, insertion into reverse micelles generates a more folded structure, characterized by the presence of 20% alpha-helix. This conformation is unaffected by variations in the water content of the system (in the 2.0-22.4 w0 range). The reactivity of epsilon-amino groups of lysine residues with aqueous solutions of o-phthalaldehyde demonstrates that segments of the peptide chain are accessible to water. Similar results were obtained with the sequence involved in heme binding. In contrast, the sole tryptophan residue, Trp-117, is shielded from the aqueous solvent, as indicated by lack of reaction with N-bromosuccinimide. The invariance of the wavelength maximum emission in the fluorescence spectra as a function of w0 is consistent with this result.(ABSTRACT TRUNCATED AT 250 WORDS)
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Nicot
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Vacher
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Vincent
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Gallay
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Waks
D000818:Animals; D001921:Brain; D002417:Cattle; D002942:Circular Dichroism; D004143:Dioctyl Sulfosuccinic Acid; D007700:Kinetics; D008239:Lysine / Q000032:analysis; D008565:Membrane Proteins / Q000378:metabolism*; D008823:Micelles; D008954:Models, Biological; D004676:Myelin Basic Protein / Q000378:metabolism*; D011487:Protein Conformation; D012995:Solubility; D013050:Spectrometry, Fluorescence; D013386:Succinates; D014364:Tryptophan / Q000032:analysis
D008565:Membrane Proteins; D008823:Micelles; D004676:Myelin Basic Protein; D013386:Succinates; D004143:Dioctyl Sulfosuccinic Acid; D014364:Tryptophan; D008239:Lysine