Peptide mapping of proteins in cerebrospinal fluid utilizing a rapid preparative two-dimensional electrophoretic procedure and matrix-assisted laser desorption/ionization mass spectrometry

Pia Davidsson, Carol L. Nilsson

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)391-399
Number of pages9
JournalBiochimica et Biophysica Acta - General Subjects
Volume1473
Issue number2-3
DOIs
StatePublished - Dec 27 1999
Externally publishedYes

Fingerprint

Cerebrospinal fluid
Cerebrospinal Fluid Proteins
Peptide Mapping
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Ionization
Mass spectrometry
Desorption
Peptides
Lasers
Glycoproteins
Proteins
Brain
Apolipoprotein A-I
Isoelectric Focusing
Brain Diseases
Apolipoproteins E
Transferrin
Liquids
Gels
Purification

Keywords

  • Cerebrospinal fluid
  • Electroelution
  • Liquid phase IEF
  • Matrix-assisted laser desorption/ionization mass spectrometry
  • Peptide mapping
  • Preparative two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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abstract = "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.",
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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.

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