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Proteome studies of human cerebrospinal fluid and brain tissue using a preparative two-dimensional electrophoresis approach prior to mass spectrometry

Davidsson, P; Paulson, Linda; Hesse, C; Blennow, K and Nilsson, Carol LU (2001) In Proteomics 1(3). p.52-444
Abstract

A preparative proteomic approach, involving liquid phase isoelectric focusing (IEF) in combination with one-dimensional electrophoresis and electroelution followed by mass spectrometry and database searches, was found to be an important tool for identifying low-abundant proteins (microgram/L) in human cerebrospinal fluid (CSF) and membrane proteins in human frontal cortex. Several neuron-related proteins, such as amyloid precursor-like protein, chromogranins A and B, glial fibrillary acid protein, beta-trace, transthyretin, ubiquitin, and cystatin C, were identified in CSF. Several types of proteins were also characterized from a detergent-solubilized human frontal cortex homogenate including membrane proteins such as synaptophysin,... (More)

A preparative proteomic approach, involving liquid phase isoelectric focusing (IEF) in combination with one-dimensional electrophoresis and electroelution followed by mass spectrometry and database searches, was found to be an important tool for identifying low-abundant proteins (microgram/L) in human cerebrospinal fluid (CSF) and membrane proteins in human frontal cortex. Several neuron-related proteins, such as amyloid precursor-like protein, chromogranins A and B, glial fibrillary acid protein, beta-trace, transthyretin, ubiquitin, and cystatin C, were identified in CSF. Several types of proteins were also characterized from a detergent-solubilized human frontal cortex homogenate including membrane proteins such as synaptophysin, syntaxin and Na+/K+ ATPase. One-third of the identified proteins have not previously been identified in human CSF or human frontal cortex using proteomic techniques. The absence of these proteins in two-dimensional electrophoresis maps might be due to insufficient amounts or low solubility. The advantages of using preparative liquid phase electrophoretic separations for identifying proteins from complex biological mixtures are speed of analysis, high loadability in the IEF separation, nondiscrimination of membrane proteins or low abundance proteins, yielding sufficient amounts for characterization by mass spectrometry. The use of this strategy in proteome studies of CSF/brain tissue is expected to offer new perspectives in studies of the pathology of neurodegenerative diseases, and reveal new potential markers for brain disorders.

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author
publishing date
type
Contribution to journal
publication status
published
keywords
Brain Chemistry, Cerebrospinal Fluid Proteins, Databases, Protein, Electrophoresis, Gel, Two-Dimensional, Humans, Isoelectric Focusing, Nerve Tissue Proteins, Proteome, Sodium Dodecyl Sulfate, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
in
Proteomics
volume
1
issue
3
pages
9 pages
publisher
John Wiley & Sons
external identifiers
  • scopus:0035294659
ISSN
1615-9853
DOI
10.1002/1615-9861(200103)1:3<444::AID-PROT444>3.0.CO;2-Q
language
English
LU publication?
no
id
42497d14-1d57-47cd-98f3-49dbc56d7a25
date added to LUP
2017-05-16 10:42:28
date last changed
2018-01-07 12:03:44
@article{42497d14-1d57-47cd-98f3-49dbc56d7a25,
  abstract     = {<p>A preparative proteomic approach, involving liquid phase isoelectric focusing (IEF) in combination with one-dimensional electrophoresis and electroelution followed by mass spectrometry and database searches, was found to be an important tool for identifying low-abundant proteins (microgram/L) in human cerebrospinal fluid (CSF) and membrane proteins in human frontal cortex. Several neuron-related proteins, such as amyloid precursor-like protein, chromogranins A and B, glial fibrillary acid protein, beta-trace, transthyretin, ubiquitin, and cystatin C, were identified in CSF. Several types of proteins were also characterized from a detergent-solubilized human frontal cortex homogenate including membrane proteins such as synaptophysin, syntaxin and Na+/K+ ATPase. One-third of the identified proteins have not previously been identified in human CSF or human frontal cortex using proteomic techniques. The absence of these proteins in two-dimensional electrophoresis maps might be due to insufficient amounts or low solubility. The advantages of using preparative liquid phase electrophoretic separations for identifying proteins from complex biological mixtures are speed of analysis, high loadability in the IEF separation, nondiscrimination of membrane proteins or low abundance proteins, yielding sufficient amounts for characterization by mass spectrometry. The use of this strategy in proteome studies of CSF/brain tissue is expected to offer new perspectives in studies of the pathology of neurodegenerative diseases, and reveal new potential markers for brain disorders.</p>},
  author       = {Davidsson, P and Paulson, Linda and Hesse, C and Blennow, K and Nilsson, Carol},
  issn         = {1615-9853},
  keyword      = {Brain Chemistry,Cerebrospinal Fluid Proteins,Databases, Protein,Electrophoresis, Gel, Two-Dimensional,Humans,Isoelectric Focusing,Nerve Tissue Proteins,Proteome,Sodium Dodecyl Sulfate,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization},
  language     = {eng},
  number       = {3},
  pages        = {52--444},
  publisher    = {John Wiley & Sons},
  series       = {Proteomics},
  title        = {Proteome studies of human cerebrospinal fluid and brain tissue using a preparative two-dimensional electrophoresis approach prior to mass spectrometry},
  url          = {http://dx.doi.org/10.1002/1615-9861(200103)1:3<444::AID-PROT444>3.0.CO;2-Q},
  volume       = {1},
  year         = {2001},
}