Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS
AuthorLothian, A; Roberts, BR
Source TitleJournal of Visualized Experiments
PublisherJOURNAL OF VISUALIZED EXPERIMENTS
University of Melbourne Author/sRoberts, Blaine
AffiliationFlorey Department of Neuroscience and Mental Health
Document TypeJournal Article
CitationsLothian, A. & Roberts, B. R. (2016). Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2016 (110), https://doi.org/10.3791/53737.
Access StatusOpen Access
Metals are essential for protein function as cofactors to catalyze chemical reactions. Disruption of metal homeostasis is implicated in a number of diseases including Alzheimer's and Parkinson's disease, but the exact role these metals play is yet to be fully elucidated. Identification of metalloproteins encounters many challenges and difficulties. Here we report an approach that allows metalloproteins in complex samples to be quantified. This is achieved using size exclusion chromatography coupled with inductively coupled plasma - mass spectrometry (SEC-ICP-MS). Using six known metalloproteins, the size exclusion column can be calibrated and the respective trace elements (iron, copper, zinc, cobalt, iodine) can be used for quantification. SEC-ICP-MS traces of human brain and plasma are presented. The use of these metalloprotein standards provides the means to quantitatively compare metalloprotein abundances between biological samples. This technique is poised to help shed light on the role of metalloproteins in neurodegenerative disease as well as other diseases where imbalances in trace elements are implicated.
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