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Localized intracellular proteome analysis using a spatially restricted enzymatic tag

Localized intracellular proteome analysis could be used as a discovery tool to map the protein components of organelles in diseased cells. To label proteins present in a specific cellular compartment, ascorbate peroxidase enzyme was linked to a mitochondrial matrix localization sequence and expressed in cultured human cells. Ascorbate peroxidase-mediated biotinylation followed by mass spectrometry analysis identified 495 mitochondrial matrix proteins, 94% of which had previously been associated with mitochondria. Next steps include using the approach to study the composition of additional organelles. Applications to disease could include characterizing the proteome of mitochondria in genetic disorders linked to mitochondrial dysfunction.

SciBX 6(5); doi:10.1038/scibx.2013.123
Published online Feb. 7, 2013

Patent application filed; available for licensing

Rhee, H.-W. et al. Science; published online Jan. 31, 2013;
doi:10.1126/science.1230593
Contact: Alice Y. Ting, Massachusetts Institute of Technology, Cambridge, Mass.
e-mail:
ating@mit.edu