Conservation of topology, but not conformation, of the proteolipid proteins of the myelin sheath.
The proteolipid protein gene products DM-20 and PLP are adhesive intrinsic membrane proteins that make up >/=50% of the protein in myelin and serve to stabilize compact myelin sheaths at the extracellular surfaces of apposed membrane lamellae. To identify which domains of DM-20 and PLP are positioned topologically in the extracellular space to participate in adhesion, we engineered N-glycosylation consensus sites into the hydrophilic segments and determined the extent of glycosylation. In addition, we assessed the presence of two translocation stop-transfer signals and, finally, mapped the extracellular and cytoplasmic dispositions of four antibody epitopes. We find that the topologies of DM-20 and PLP are identical, with both proteins possessing four transmembrane domains and N and C termini exposed to the cytoplasm. Consistent with this notion, DM-20 and PLP contain within their N- and C-terminal halves independent stop-transfer signals for insertion into the bilayer of the rough endoplasmic reticulum during de novo synthesis. Surprisingly, the conformation (as opposed to topology) of DM-20 and PLP may differ, which has been inferred from the divergent effects that many missense mutations have on the intracellular trafficking of these two isoforms. The 35 amino acid cytoplasmic peptide in PLP, which distinguishes this protein from DM-20, imparts a sensitivity to mutations in extracellular domains. This peptide may normally function during myelinogenesis to detect conformational changes originating across the bilayer from extracellular PLP interactions in trans and trigger intracellular events such as membrane compaction in the cytoplasmic compartment.
Gow
A
A
Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029-6574, USA.
Gragerov
A
A
Gard
A
A
Colman
D R
DR
Lazzarini
R A
RA
D000911:Antibodies, Monoclonal; D001059:Apoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004251:DNA Transposable Elements; D005455:Fluorescent Antibody Technique; D006801:Humans; D008861:Microsomes / Q000378:metabolism; D009154:Mutation; D018991:Myelin Proteolipid Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010449:Peptide Mapping; D017354:Point Mutation; D011487:Protein Conformation; D015398:Signal Transduction; D014162:Transfection
D000911:Antibodies, Monoclonal; D001059:Apoproteins; D004251:DNA Transposable Elements; D018991:Myelin Proteolipid Protein; D010446:Peptide Fragments