Identification of proteins from human cerebrospinal fluid, separated by two-dimensional polyacrylamide gel electrophoresis.
The aim of this work is to display the protein composition of the cerebrospinal fluid by two-dimensional (2-D) gel electrophoresis and identify it using different mass spectrometric techniques. This will enable us to present an overview of the proteins in human cerebrospinal fluid. The comparison of 2-D gels will help us to analyze the normal protein variability in healthy persons and specific protein variations in patients with different neurological diseases (e.g., morbus Alzheimer, chorea Huntington). However, it is not possible to carry out 2-D gel electrophoresis directly with human cerebrospinal fluid due to the high amount of salts, sugars and lipids present. In addition, the total amount of protein is only as high as 0.3-0.7 microg/microL. Therefore, concentration and desalting steps using precipitation and ultrafiltration are necessary. To date we have been able to identify more than 65 spots from 2-D gels using matrix assisted laser desorption/ionization-mass spectrometry and electrospray ionization-mass spectrometry.
Sickmann
A
A
Institut für Physiologische Chemie, Ruhr-Universität Bochum, Germany. albert.sickmann@ruhr-uni-bochum.de
Dormeyer
W
W
Wortelkamp
S
S
Woitalla
D
D
Kuhn
W
W
Meyer
H E
HE
D000595:Amino Acid Sequence; D002556:Cerebrospinal Fluid Proteins / Q000032:analysis* / Q000302:isolation & purification; D011232:Chemical Precipitation; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007525:Isoelectric Focusing; D008715:Methionine / Q000737:chemistry; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D012016:Reference Values; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013382:Subtraction Technique; D014462:Ultrafiltration
D002556:Cerebrospinal Fluid Proteins; D008715:Methionine