Interaction of glycosylated human myelin basic protein with lipid bilayers.
Myelin basic protein (MBP), isolated from normal human myelin, was glycosylated with UDP-N-acetyl-D-galactosamine and a glycosyltransferase isolated from porcine submaxillary glands. MBP containing 0.85 mol of N-acetyl-D-galactosamine per mole of protein was oxidized at carbon 6 by galactose oxidase and complexed with a spin-label, Tempoamine, in order to study its interactions with lipids. When the spin-labeled MBP was reacted with lipid vesicles consisting of DSPG, DPPG, and DMPG, most of the spin-label was motionally restricted in the gel phase, with a correlation time greater than 10(-8)s. The motion increased with increasing temperature and was sensitive to the lipid phase transition. Interaction with the gel phase of DPPA caused much less motional restriction of the probe. However, melting of the lipid allowed increased interaction and motional restriction of the probe, which was only partially reversed on cooling back to the gel phase. The motional restriction of the probe in these lipids is attributed to its penetration partway into the lipid bilayer in both the gel and liquid-crystalline phases. The fact that the probe bound to the protein can penetrate partway into the bilayer suggests that other hydrophobic side chains and residues of the protein can similarly penetrate into the bilayer. Additional evidence for penetration was provided by digestion of the lipid-bound protein with endoproteinase Lys-C. When nonglycosylated and glycosylated MBP in solution was treated with Lys-C, extensive digestion occurred. A single radioactive peptide which eluted at 25 min was identified as residues 92-105.(ABSTRACT TRUNCATED AT 250 WORDS)
Persaud
R
R
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
Boggs
J M
JM
Wood
D D
DD
Moscarello
M A
MA
D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D008051:Lipid Bilayers / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000378:metabolism*; D010712:Phosphatidic Acids / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D011487:Protein Conformation; D013113:Spin Labels
D008051:Lipid Bilayers; D004676:Myelin Basic Protein; D010712:Phosphatidic Acids; D010715:Phosphatidylglycerols; D013113:Spin Labels; C030345:1,2-dipalmitoylphosphatidylglycerol