Photolabeling of myelin basic protein in lipid vesicles with the hydrophobic reagent 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine.
The hydrophobic photolabel 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine([125I]TID) was used to label myelin basic protein or polylysine in aqueous solution and bound to lipid vesicles of different composition. Although myelin basic protein is a water soluble protein which binds electrostatically only to acidic lipids, unlike polylysine it has several short hydrophobic regions. Myelin basic protein was labeled to a significant extent by TID when in aqueous solution indicating that it has a hydrophobic site which can bind the reagent. However, myelin basic protein was labeled 2-4-times more when bound to the acidic lipids phosphatidylglycerol, phosphatidylserine, phosphatidic acid, and cerebroside sulfate than when bound to phosphatidylethanolamine, or when in solution in the presence of phosphatidylcholine vesicles. It was labeled 5-7-times more than polylysine bound to acidic lipids. These results suggest that when myelin basic protein is bound to acidic lipids, it is labeled from the lipid bilayer rather than from the aqueous phase. However, this conclusion is not unequivocal because of the possibility of changes in the protein conformation or degree of aggregation upon binding to lipid. Within this limitation the results are consistent with, but do not prove, the concept that some of its hydrophobic residues penetrate partway into the lipid bilayer. However, it is likely that most of the protein is on the surface of the bilayer with its basic residues bound electrostatically to the lipid head groups.
D000818:Animals; D001389:Azirines / Q000378:metabolism*; D002554:Cerebrosides / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D004676:Myelin Basic Protein / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D010777:Photochemistry; D011107:Polylysine / Q000378:metabolism; D012995:Solubility; D014961:X-Ray Diffraction
D001389:Azirines; D002554:Cerebrosides; D004676:Myelin Basic Protein; D010743:Phospholipids; D011107:Polylysine; C033174:3-(trifluoromethyl)-3-(3-iodophenyl)diazirine