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Oxidative metabolism of dopamine: a colour reaction from human midbrain analysed by mass spectrometry

De Iuliis, A; Arrigoni, Giorgio LU ; Andersson, L; Zambenedetti, P; Burlina, A; James, Peter LU ; Arslan, P and Vianello, F (2008) In Biochimica et Biophysica Acta - Proteins and Proteomics 1784. p.1687-1693
Abstract
In order to identify the protein responsible for a dopamine peroxidizing activity, previously described in human normal and parkinsonian substantia nigra by our group, we developed non-denaturing polyacrylamide gel electrophoresis conditions, mimicking the characteristic colour in vitro reaction, resulting from cyclic oxidation of dopamine (DA). After separating protein mixtures from human normal midbrain homogenates on two sets of identical native gels, one gel set was subjected to specific activity staining by using DA and hydrogen peroxide. An activity red/orange band appeared in midbrain tissue lanes, similarly to the lane where commercial horseradish peroxidase (HRP) was present as control of peroxidative activity. The second set of... (More)
In order to identify the protein responsible for a dopamine peroxidizing activity, previously described in human normal and parkinsonian substantia nigra by our group, we developed non-denaturing polyacrylamide gel electrophoresis conditions, mimicking the characteristic colour in vitro reaction, resulting from cyclic oxidation of dopamine (DA). After separating protein mixtures from human normal midbrain homogenates on two sets of identical native gels, one gel set was subjected to specific activity staining by using DA and hydrogen peroxide. An activity red/orange band appeared in midbrain tissue lanes, similarly to the lane where commercial horseradish peroxidase (HRP) was present as control of peroxidative activity. The second set of gels, stained with Coomassie Blue, showed other, not enzymatically active protein bands. Mass spectrometry analysis of the bands containing the activity and the corresponding Coomassie Blue bands revealed the presence of proteins that may play a role in neurodegenerative disease, highlighting a possible functional link among dopamine/dopaminochrome redox cycle and protein metabolism. (Less)
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author
organization
publishing date
type
Contribution to specialist publication or newspaper
publication status
published
subject
categories
Popular Science
in
Biochimica et Biophysica Acta - Proteins and Proteomics
volume
1784
pages
1687 - 1693
external identifiers
  • PMID:18675943
  • WOS:000261019000025
  • Scopus:54049089936
ISSN
1570-9639
DOI
10.1016/j.bbapap.2008.07.002
language
English
LU publication?
yes
id
eca87be0-f7fa-474e-a1e5-8de2cbfa3eba (old id 1244429)
date added to LUP
2008-10-07 11:28:42
date last changed
2016-10-13 04:32:36
@misc{eca87be0-f7fa-474e-a1e5-8de2cbfa3eba,
  abstract     = {In order to identify the protein responsible for a dopamine peroxidizing activity, previously described in human normal and parkinsonian substantia nigra by our group, we developed non-denaturing polyacrylamide gel electrophoresis conditions, mimicking the characteristic colour in vitro reaction, resulting from cyclic oxidation of dopamine (DA). After separating protein mixtures from human normal midbrain homogenates on two sets of identical native gels, one gel set was subjected to specific activity staining by using DA and hydrogen peroxide. An activity red/orange band appeared in midbrain tissue lanes, similarly to the lane where commercial horseradish peroxidase (HRP) was present as control of peroxidative activity. The second set of gels, stained with Coomassie Blue, showed other, not enzymatically active protein bands. Mass spectrometry analysis of the bands containing the activity and the corresponding Coomassie Blue bands revealed the presence of proteins that may play a role in neurodegenerative disease, highlighting a possible functional link among dopamine/dopaminochrome redox cycle and protein metabolism.},
  author       = {De Iuliis, A and Arrigoni, Giorgio and Andersson, L and Zambenedetti, P and Burlina, A and James, Peter and Arslan, P and Vianello, F},
  issn         = {1570-9639},
  language     = {eng},
  pages        = {1687--1693},
  series       = {Biochimica et Biophysica Acta - Proteins and Proteomics},
  title        = {Oxidative metabolism of dopamine: a colour reaction from human midbrain analysed by mass spectrometry},
  url          = {http://dx.doi.org/10.1016/j.bbapap.2008.07.002},
  volume       = {1784},
  year         = {2008},
}