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Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins
Transport of LDL-derived cholesterol from lysosomes into the cytoplasm requires NPC1 protein; NPC1L1 mediates uptake of dietary cholesterol. We introduced single disulfide bonds into NPC1 and NPC1L1 to explore the importance of inter-domain dynamics in cholesterol transport. Using a sensitive method...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7228765/ https://www.ncbi.nlm.nih.gov/pubmed/32410728 http://dx.doi.org/10.7554/eLife.57089 |
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author | Saha, Piyali Shumate, Justin L Caldwell, Jenna G Elghobashi-Meinhardt, Nadia Lu, Albert Zhang, Lichao Olsson, Niclas E Elias, Joshua E Pfeffer, Suzanne R |
author_facet | Saha, Piyali Shumate, Justin L Caldwell, Jenna G Elghobashi-Meinhardt, Nadia Lu, Albert Zhang, Lichao Olsson, Niclas E Elias, Joshua E Pfeffer, Suzanne R |
author_sort | Saha, Piyali |
collection | PubMed |
description | Transport of LDL-derived cholesterol from lysosomes into the cytoplasm requires NPC1 protein; NPC1L1 mediates uptake of dietary cholesterol. We introduced single disulfide bonds into NPC1 and NPC1L1 to explore the importance of inter-domain dynamics in cholesterol transport. Using a sensitive method to monitor lysosomal cholesterol efflux, we found that NPC1’s N-terminal domain need not release from the rest of the protein for efficient cholesterol export. Either introducing single disulfide bonds to constrain lumenal/extracellular domains or shortening a cytoplasmic loop abolishes transport activity by both NPC1 and NPC1L1. The widely prescribed cholesterol uptake inhibitor, ezetimibe, blocks NPC1L1; we show that residues that lie at the interface between NPC1L1's three extracellular domains comprise the drug’s binding site. These data support a model in which cholesterol passes through the cores of NPC1/NPC1L1 proteins; concerted movement of various domains is needed for transfer and ezetimibe blocks transport by binding to multiple domains simultaneously. |
format | Online Article Text |
id | pubmed-7228765 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-72287652020-05-18 Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins Saha, Piyali Shumate, Justin L Caldwell, Jenna G Elghobashi-Meinhardt, Nadia Lu, Albert Zhang, Lichao Olsson, Niclas E Elias, Joshua E Pfeffer, Suzanne R eLife Biochemistry and Chemical Biology Transport of LDL-derived cholesterol from lysosomes into the cytoplasm requires NPC1 protein; NPC1L1 mediates uptake of dietary cholesterol. We introduced single disulfide bonds into NPC1 and NPC1L1 to explore the importance of inter-domain dynamics in cholesterol transport. Using a sensitive method to monitor lysosomal cholesterol efflux, we found that NPC1’s N-terminal domain need not release from the rest of the protein for efficient cholesterol export. Either introducing single disulfide bonds to constrain lumenal/extracellular domains or shortening a cytoplasmic loop abolishes transport activity by both NPC1 and NPC1L1. The widely prescribed cholesterol uptake inhibitor, ezetimibe, blocks NPC1L1; we show that residues that lie at the interface between NPC1L1's three extracellular domains comprise the drug’s binding site. These data support a model in which cholesterol passes through the cores of NPC1/NPC1L1 proteins; concerted movement of various domains is needed for transfer and ezetimibe blocks transport by binding to multiple domains simultaneously. eLife Sciences Publications, Ltd 2020-05-15 /pmc/articles/PMC7228765/ /pubmed/32410728 http://dx.doi.org/10.7554/eLife.57089 Text en © 2020, Saha et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Biochemistry and Chemical Biology Saha, Piyali Shumate, Justin L Caldwell, Jenna G Elghobashi-Meinhardt, Nadia Lu, Albert Zhang, Lichao Olsson, Niclas E Elias, Joshua E Pfeffer, Suzanne R Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title | Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title_full | Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title_fullStr | Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title_full_unstemmed | Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title_short | Inter-domain dynamics drive cholesterol transport by NPC1 and NPC1L1 proteins |
title_sort | inter-domain dynamics drive cholesterol transport by npc1 and npc1l1 proteins |
topic | Biochemistry and Chemical Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7228765/ https://www.ncbi.nlm.nih.gov/pubmed/32410728 http://dx.doi.org/10.7554/eLife.57089 |
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