MassIVE MSV000079096

Partial Public PXD002017

Proteomic Mapping of Mitochondria in Living Cells via Spatially Restricted Enzymatic Tagging

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Description

Rhee HW, Zou P, Udeshi ND, Martell JD, Mootha VK, Carr SA,Ting AY. Science, 2013, 339, 1328-1331. doi: 10.1126/science.1230593. Microscopy and mass spectrometry (MS) are complementary techniques: the former provides spatiotemporal information in living cells, but only for a handful of recombinant proteins, while the latter can detect thousands of endogenous proteins simultaneously, but only in lysed samples. Here we introduce technology that combines these strengths by offering spatially- and temporally resolved proteomic maps of endogenous proteins within living cells. The method relies on a genetically-targetable peroxidase enzyme that biotinylates nearby proteins, which are subsequently purified and identified by MS. We used this approach to identify 495 proteins within the human mitochondrial matrix, including 31 not previously linked to mitochondria. The labeling was exceptionally specific and distinguished between inner membrane proteins facing the matrix versus the intermembrane space (IMS). Several proteins previously thought to reside in the IMS or outer membrane, including protoporphyrinogen oxidase, were reassigned to the matrix. The specificity of live-cell peroxidase-mediated proteomic mapping combined with its ease of use offers biologists a powerful tool for understanding the molecular composition of living cells. [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: SILAC ; mitochondria ; APEX

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Principal Investigators:
(in alphabetical order) 		Steven A. Carr
Submitting User: clauser
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