TY - JOUR
T1 - Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry
AU - Davidsson, Pia
AU - Nilsson, Carol L.
N1 - Funding Information:
This work was supported by grants from The Swedish Medical Research Council (Grants 12769, 13121); Foundation for Strategic Research (Infection and Vaccinology), Alzheimerfonden, Lund, Sweden; Bohuslandstingets FoU fond, Sweden; Stiftelsen Gamla Tjännarinnor, Stochholm, Sweden, Swedish Medical Society, Stiftelsen Handlanden Hjalmar Svenssons Forskningsfond, Göteborg, Sweden; Loo and Hans Osterman fond, Stockholm, Sweden; Lundbecksstiftelsen, Lund, Sweden; Wilhelm och Martina Lundgrens Stiftelse, Göteborg, Sweden, Magnus Bergvalls Stiftelse, Stockholm, Sweden; Thurings Stiftelse, Stockholm, Sweden, Åke Wibergs Stiftelse, Stockholm, Sweden. The authors would like to thank Thomas Larsson for participating in helpful discussions.
PY - 1999/12/27
Y1 - 1999/12/27
N2 - A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.
AB - A quick two-step procedure involving liquid phase isoelectric focusing in the Rotofor cell in combination with electroelution in the Mini whole cell gel eluter has been used for purification of proteins from human cerebrospinal fluid (CSF). Fractions, each highly enriched in a single protein band and virtually free of other proteins, were selected for characterization by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOFMS). Six CSF proteins, transferrin, α1-acid-glycoprotein, Zn-α2-glycoprotein, apolipoprotein A1, apolipoprotein E and β-trace were identified by MALDI-TOFMS analysis of the tryptic digests. These results demonstrate that the combination of liquid phase IEF and electroelution is a rapid preparative two-dimensional separation which can provide single proteins of high purity, in yields sufficient for characterization by MALDI-TOFMS. Characterization of such brain-specific proteins in CSF will be useful in the investigation of the pathophysiology of different brain disorders. Copyright (C) 1999 Elsevier Science B.V.
KW - Cerebrospinal fluid
KW - Electroelution
KW - Liquid phase IEF
KW - Matrix-assisted laser desorption/ionization mass spectrometry
KW - Peptide mapping
KW - Preparative two-dimensional gel electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=0032713887&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032713887&partnerID=8YFLogxK
U2 - 10.1016/S0304-4165(99)00197-X
DO - 10.1016/S0304-4165(99)00197-X
M3 - Article
C2 - 10594376
AN - SCOPUS:0032713887
SN - 0304-4165
VL - 1473
SP - 391
EP - 399
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
IS - 2-3
ER -