An immunologic probe for a defined region of the myelin proteolipid.
Antiserum has been prepared against an isolated polypeptide fragment, designated BPS4, which comprises residues 181-211 of the bovine myelin proteolipid. The antiserum recognizes the intact bovine proteolipid protein but not several other polypeptide fragments within the molecule, nor the myelin basic protein, thus demonstrating specificity of the antiserum. In a competitive enzyme-linked immunosorbent assay, both the major proteolipid and the DM 20 bands observed on sodium dodecyl sulfate-polyacrylamide gels reacted equally well with the antiserum, indicating that the BPS4 segment is present in both molecular species. The rat myelin proteolipid protein cross-reacted with antiserum against the intact bovine protein but showed minimal cross-reactivity with the antiserum against the bovine BPS4 fragment. This was demonstrated in parallel experiments using three types of preparations, namely, sodium dodecyl sulfate-solubilized myelin, delipidated myelin, and isolated proteolipid apoprotein. The difference between the bovine and rat proteins, which presumably reflects amino acid sequence differences, is thus detectable by the antiserum against the polypeptide fragment but not by the antiserum against the intact protein. Isolated bovine myelin membranes did not bind the antiserum in the absence of detergent or without delipidation. On the other hand, in vesicles reconstituted with the intact bovine apoprotein, the BPS4 segment was oriented on the exterior face of the liposome where it was capable of binding antibody and was susceptible to Pronase digestion.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000906:Antibodies; D002417:Cattle; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D009186:Myelin Sheath / Q000032:analysis*; D010446:Peptide Fragments / Q000276:immunology*; D011510:Proteolipids / Q000032:analysis*; D051381:Rats; D013045:Species Specificity
D000596:Amino Acids; D000906:Antibodies; D010446:Peptide Fragments; D011510:Proteolipids