ABSTRACT
Small and preparative gel filtration columns were studied for high pressure liquid chromatography of myelin proteins in sodium dodecyl sulfate. The preparative column proved useful for isolating and purifying proteolipid protein almost free (0.3-0.5%) of myelin basic protein as demonstrated by SDS-PAGE, MBP RIA, and immunoblotting. The small columns were not as useful as SDS-PAGE for analytical purposes.
Subject(s)
Myelin Proteins/isolation & purification , Myelin Sheath/analysis , Proteolipids/isolation & purification , Chromatography, Gel , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Humans , Radioimmunoassay , Sodium Dodecyl SulfateABSTRACT
The myelin specific proteins, myelin basic protein (MBP) and myelin proteolipid protein (PLP) were quantitated by radioimmunoassay (RIA) and the activity of the enzyme 2',3'-cyclic 3' phosphohydrolase (CNP) measured, in 27 regions of normal brain and spinal cord. Varying regional concentrations for each protein and regional variations for protein ratios were noted, supporting the concept of a varying chemical composition for myelin throughout the central nervous system (CNS). Variation was also noted among myelin subfractions from a single region. Regions with special sensitivity to the multiple sclerosis process had relatively lower proportions of CNP in several, but not all cases.
Subject(s)
2',3'-Cyclic-Nucleotide Phosphodiesterases/analysis , Brain Chemistry , Myelin Basic Protein/analysis , Myelin Proteins/analysis , Phosphoric Diester Hydrolases/analysis , Spinal Cord/analysis , Adult , Brain/enzymology , Humans , Middle Aged , Myelin Proteolipid Protein , Radioimmunoassay/methods , Spinal Cord/enzymology , Tissue DistributionABSTRACT
Using radioimmunoassays (RIA) for the myelin specific proteins, myelin proteolipid protein (PLP) and myelin basic protein (MBP) and an enzyme assay for the activity of the myelin marker enzyme 2'3' cyclic-3' phosphohydrolase (CNPase), we have studied plaque, periplaque and normal appearing white matter (NAWM) tissue. We found that all three mylein proteins are decreased in all regions, including NAWM, of MS brain, with a decreasing gradient from NAWM to perregion. Surprisingly, when the ratios of the proteins were calculated, MBP activity, although decreased was found to be relatively preserved.
Subject(s)
2',3'-Cyclic-Nucleotide Phosphodiesterases/analysis , Brain Chemistry , Multiple Sclerosis/metabolism , Myelin Basic Protein/analysis , Myelin Proteins/analysis , Phosphoric Diester Hydrolases/analysis , Brain/enzymology , Humans , Myelin Proteolipid ProteinABSTRACT
Two hundred seventy-eight samples of cerebrospinal fluid and 273 samples of sera from a variety of neurological patients were examined by radioimmunoassay for myelin proteolipid protein-like reactivity as an index of structural damage to myelin. Groups of patients with active central nervous system damage could be distinguished easily, although there was a small percentage of false positive reactions. The reactivity was most marked in the demyelinative, encephalitic, and stroke categories. A radioimmunoassay for proteolipid protein of proper sensitivity and specificity may prove to be a useful objective clinical measure of the presence and degree of disease activity in multiple sclerosis and other diseases.
