Quantification of stable isotope tracers after metabolic labeling provides a snapshot of the dynamic state of living cells and tissue. A form of imaging mass spectrometry quantifies isotope ratios with a lateral resolution <50 nm, using a methodology that we refer to as multi-isotope imaging mass spectrometry (MIMS). Despite lateral resolution exceeding diffraction-limited light microscopy, lack of contrast has largely limited use of MIMS to large or specialized subcellular structures, such as the nucleus and stereocilia. In this study, we repurpose the engineered peroxidase APEX2 as the first genetically encoded marker for MIMS. Coupling APEX2 labeling of lysosomes and metabolic labeling of protein, we identify that individual lysosomes exhibit substantial heterogeneity in protein age, which is lost in iPSC-derived neurons lacking the lysosomal protein progranulin. This study expands the practical use of MIMS for cell biology by enabling measurements of metabolic function from stable isotope labeling within individual organelles in situ.
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November 12 2019
Coupling APEX labeling to imaging mass spectrometry of single organelles reveals heterogeneity in lysosomal protein turnover
Derek P. Narendra
,
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
Correspondence to Derek P. Narendra: derek.narendra@nih.gov
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Christelle Guillermier
,
Christelle Guillermier
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
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Frank Gyngard
,
Frank Gyngard
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
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Xiaoping Huang
,
Xiaoping Huang
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
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Michael E. Ward
,
Michael E. Ward
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
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Matthew L. Steinhauser
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
Matthew L. Steinhauser: msteinhauser@pitt.edu
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Derek P. Narendra
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
Christelle Guillermier
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
Frank Gyngard
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
Xiaoping Huang
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
Michael E. Ward
1
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD
Matthew L. Steinhauser
2
Harvard Medical School, Boston, MA
3
Center for NanoImaging, Cambridge, MA
4
Department of Medicine, Division of Genetics, Brigham and Women’s Hospital, Boston, MA
Correspondence to Derek P. Narendra: derek.narendra@nih.gov
Matthew L. Steinhauser: msteinhauser@pitt.edu
J Cell Biol (2020) 219 (1): e201901097.
Article history
Received:
January 17 2019
Revision Received:
September 13 2019
Accepted:
October 08 2019
Citation
Derek P. Narendra, Christelle Guillermier, Frank Gyngard, Xiaoping Huang, Michael E. Ward, Matthew L. Steinhauser; Coupling APEX labeling to imaging mass spectrometry of single organelles reveals heterogeneity in lysosomal protein turnover. J Cell Biol 6 January 2020; 219 (1): e201901097. doi: https://doi.org/10.1083/jcb.201901097
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