Study of human cerebrospinal fluid proteins by size exclusion-high performance liquid chromatography and two-dimensional gel electrophoresis.
Cerebrospinal fluid (CSF) proteins were separated into three main fractions by size exclusion-high performance liquid chromatography (SE-HPLC). Subsequent analysis of each fraction by two-dimensional gel electrophoresis (2-DE) facilitated the detection of trace components in CSF and additionally provided more information about the native properties of various proteins. Certain proteins are present in a polymeric form and appear in the high molecular weight SE-HPLC fraction. In the middle molecular weight SE-HPLC fraction we found a CSF-specific transthyretin-related protein by immunoblotting with polyclonal antibodies to transthyretin. Possible interpolypeptide disulfide bonds of such polymeric proteins were studied using a nonreducing 2-DE system. This procedure revealed that all apolipoprotein E monomers in CSF, which are synthesized in astrocytes, are linked by disulfide bonds. In the CSF from a patient with clinically definite multiple sclerosis (MS), novel proteins appeared in the high molecular weight SE-HPLC fraction, which are obscured by other proteins if total CSF is analyzed.
Institute of Clinical Chemistry, University Hospital, Zürich, Switzerland.
F
F
Wiederkehr
M R
MR
Büeler
M
M
Wacker
D J
DJ
Vonderschmitt
D002556:Cerebrospinal Fluid Proteins / Q000302:isolation & purification*; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D004220:Disulfides / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D009422:Nervous System Diseases / Q000134:cerebrospinal fluid; D010316:Particle Size; D010449:Peptide Mapping
D002556:Cerebrospinal Fluid Proteins; D004220:Disulfides