Oxidative metabolism of dopamine: A colour reaction from human midbrain analysed by mass spectrometry
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- 作者：Angela De Iuliis ; Giorgio Arrigoni ; Liselotte Andersson ; Pamela Zambenedetti ; Aless ; ro Burlina ; Peter James ; Paola Arslan ; Fabio Vianello
- 关键词：Dopamine ; Dopaminochrome ; Dopamine peroxidizing activity ; Human brain ; Parkinson's disease ; Mass spectrometry
- 刊名：Biochimica et Biophysica Acta - Proteins and Proteomics
- 出版年：2008
- 出版时间：November 2008
- 年：2008
- 卷：1784
- 期：11
- 页码：1687-1693
- 全文大小：271 K
文摘
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.