Analysis of Trace Elements and Metalloproteins in Fractionated Human Brain Samples Using Size Exclusion Inductively Coupled Mass Spectrometry
To understand the role of trace elements in cellular function a precise mechanistic detail of their subcellular location and binding partners is required. This can be achieved by cellular fractionation and liquid chromatography coupled to inductively coupled plasma mass spectrometry (LC-ICP-MS). The increasing use of size exclusion coupled to ICP-MS allows for the specific investigation of metalloproteins. This chapter details a general cellular fractionation procedure that separates cells and tissue into three components; (1) the soluble cytosolic and extracellular component; (2) membrane proteins and vesicular cargo; and (3) highly insoluble proteins found in pathological inclusions such as amyloid plaque, neurofibrillary tangles (NFTs) and Lewy bodies. We also detail the methods to conduct a metalloproteomic analysis of soluble proteins found in biological samples with human brain as an example.
Key wordsTrace elements Metalloprotein Mass spectrometry Proteomics Tissue fractionation ICP-MS
- 4.Williams JR, Trias E, Beilby PR, Lopez NI, Labut EM, Bradford CS, Roberts BR, McAllum EJ, Crouch PJ, Rhoads TW, Pereira C, Son M, Elliott JL, Franco MC, Estevez AG, Barbeito L, Beckman JS (2016) Copper delivery to the CNS by CuATSM effectively treats motor neuron disease in SOD(G93A) mice co-expressing the copper-chaperone-for-SOD. Neurobiol Dis 89:1–9CrossRefPubMedGoogle Scholar
- 5.Roberts BR, Lim NK, McAllum EJ, Donnelly PS, Hare DJ, Doble PA, Turner BJ, Price KA, Lim SC, Paterson BM, Hickey JL, Rhoads TW, Williams JR, Kanninen KM, Hung LW, Liddell JR, Grubman A, Monty JF, Llanos RM, Kramer DR, Mercer JF, Bush AI, Masters CL, Duce JA, Li QX, Beckman JS, Barnham KJ, White AR, Crouch PJ (2014) Oral treatment with Cu(II)(atsm) increases mutant SOD1 in vivo but protects motor neurons and improves the phenotype of a transgenic mouse model of amyotrophic lateral sclerosis. J Neurosci 34:8021–8031CrossRefPubMedGoogle Scholar