Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes
The lysosome degrades and recycles macromolecules, signals to the cytosol and nucleus, and is implicated in many diseases. Here, we describe a method for the rapid isolation of mammalian lysosomes and use it to quantitatively profile lysosomal metabolites under various cell states. Under nutrient-re...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Published: |
American Association for the Advancement of Science (AAAS)
2019
|
| Online Access: | http://hdl.handle.net/1721.1/119871 https://orcid.org/0000-0003-4642-3706 https://orcid.org/0000-0002-1446-7256 |
| _version_ | 1826210194590269440 |
|---|---|
| author | Chan, Sze Ham Freinkman, Elizaveta Abu-Remaileh, Monther Wyant, Gregory Andrew Kim, Choah Laqtom, Nouf N Abbasi, Maria Sabatini, David |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Chan, Sze Ham Freinkman, Elizaveta Abu-Remaileh, Monther Wyant, Gregory Andrew Kim, Choah Laqtom, Nouf N Abbasi, Maria Sabatini, David |
| author_sort | Chan, Sze Ham |
| collection | MIT |
| description | The lysosome degrades and recycles macromolecules, signals to the cytosol and nucleus, and is implicated in many diseases. Here, we describe a method for the rapid isolation of mammalian lysosomes and use it to quantitatively profile lysosomal metabolites under various cell states. Under nutrient-replete conditions, many lysosomal amino acids are in rapid exchange with those in the cytosol. Loss of lysosomal acidification through inhibition of the vacuolar H+–adenosine triphosphatase (V-ATPase) increased the luminal concentrations of most metabolites but had no effect on those of the majority of essential amino acids. Instead, nutrient starvation regulates the lysosomal concentrations of these amino acids, an effect we traced to regulation of the mechanistic target of rapamycin (mTOR) pathway. Inhibition of mTOR strongly reduced the lysosomal efflux of most essential amino acids, converting the lysosome into a cellular depot for them. These results reveal the dynamic nature of lysosomal metabolites and that V-ATPase- and mTOR-dependent mechanisms exist for controlling lysosomal amino acid efflux. |
| first_indexed | 2024-09-23T14:45:25Z |
| format | Article |
| id | mit-1721.1/119871 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T14:45:25Z |
| publishDate | 2019 |
| publisher | American Association for the Advancement of Science (AAAS) |
| record_format | dspace |
| spelling | mit-1721.1/1198712022-09-29T10:23:35Z Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes Chan, Sze Ham Freinkman, Elizaveta Abu-Remaileh, Monther Wyant, Gregory Andrew Kim, Choah Laqtom, Nouf N Abbasi, Maria Sabatini, David Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Abu-Remaileh, Monther Wyant, Gregory Andrew Kim, Choah Laqtom, Nouf N Abbasi, Maria Sabatini, David The lysosome degrades and recycles macromolecules, signals to the cytosol and nucleus, and is implicated in many diseases. Here, we describe a method for the rapid isolation of mammalian lysosomes and use it to quantitatively profile lysosomal metabolites under various cell states. Under nutrient-replete conditions, many lysosomal amino acids are in rapid exchange with those in the cytosol. Loss of lysosomal acidification through inhibition of the vacuolar H+–adenosine triphosphatase (V-ATPase) increased the luminal concentrations of most metabolites but had no effect on those of the majority of essential amino acids. Instead, nutrient starvation regulates the lysosomal concentrations of these amino acids, an effect we traced to regulation of the mechanistic target of rapamycin (mTOR) pathway. Inhibition of mTOR strongly reduced the lysosomal efflux of most essential amino acids, converting the lysosome into a cellular depot for them. These results reveal the dynamic nature of lysosomal metabolites and that V-ATPase- and mTOR-dependent mechanisms exist for controlling lysosomal amino acid efflux. 2019-01-08T16:18:33Z 2019-01-08T16:18:33Z 2017-11 2017-05 2018-12-19T15:31:26Z Article http://purl.org/eprint/type/JournalArticle 0036-8075 1095-9203 http://hdl.handle.net/1721.1/119871 Abu-Remaileh, Monther et al. “Lysosomal Metabolomics Reveals V-ATPase- and mTOR-Dependent Regulation of Amino Acid Efflux from Lysosomes.” Science 358, 6364 (October 2017): 807–813 © 2017 American Association for the Advancement of Science https://orcid.org/0000-0003-4642-3706 https://orcid.org/0000-0002-1446-7256 http://dx.doi.org/10.1126/SCIENCE.AAN6298 Science Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf American Association for the Advancement of Science (AAAS) PMC |
| spellingShingle | Chan, Sze Ham Freinkman, Elizaveta Abu-Remaileh, Monther Wyant, Gregory Andrew Kim, Choah Laqtom, Nouf N Abbasi, Maria Sabatini, David Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title | Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title_full | Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title_fullStr | Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title_full_unstemmed | Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title_short | Lysosomal metabolomics reveals V-ATPase- and mTOR-dependent regulation of amino acid efflux from lysosomes |
| title_sort | lysosomal metabolomics reveals v atpase and mtor dependent regulation of amino acid efflux from lysosomes |
| url | http://hdl.handle.net/1721.1/119871 https://orcid.org/0000-0003-4642-3706 https://orcid.org/0000-0002-1446-7256 |
| work_keys_str_mv | AT chanszeham lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT freinkmanelizaveta lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT aburemailehmonther lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT wyantgregoryandrew lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT kimchoah lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT laqtomnoufn lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT abbasimaria lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes AT sabatinidavid lysosomalmetabolomicsrevealsvatpaseandmtordependentregulationofaminoacideffluxfromlysosomes |