Profiling the iron, copper and zinc content in primary neuron and astrocyte cultures by rapid online quantitative size exclusion chromatography-inductively coupled plasma-mass spectrometry

Dominic J Hare, Alexandra Grubman, Timothy M Ryan, Amber Lothian, Jeffrey R. Liddell, Rudolf Grimm, Toshiaki Matsude, Philip Doble, Robert A Cherny, Ashley I. Bush, Anthony R White, Colin L. Masters, Blaine R. Roberts

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32 Citations (Scopus)

Abstract

Metals often determine the chemical reactivity of the proteins to which they are bound. Each cell in the body tightly maintains a unique metalloproteomic profile, mostly dependent on function. This paper describes an analytical online flow injection quantitative size exclusion chromatography-inductively coupled plasma-mass spectrometry (SEC-ICP-MS) method, which was applied to profiling the metal-binding proteins found in primary cultures of neurons and astrocytes. This method can be conducted using similar amounts of sample to those used for Western blotting (20-150 μg protein), and has a turnaround time of -1). Total on-column Fe, Cu and Zn detection limits ranged from 0.825 ± 0.005 ng to 13.6 ± 0.7 pg. Neurons and astrocytes exhibited distinct metalloprotein profiles, featuring both ubiquitous and unique metalloprotein species. Separation and detection by SEC-ICP-MS allows appraisal of these metalloproteins in their native state, and online quantification was achieved using this relatively simple external calibration process.
Original languageEnglish
Pages (from-to)1656-1662
Number of pages7
JournalMetallomics
Volume5
Issue number12
DOIs
Publication statusPublished - Dec 2013
Externally publishedYes

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