Cargando…
Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport
ORPs are lipid-transport proteins belonging to the oxysterol-binding protein family. They facilitate the transfer of lipids between different intracellular membranes, such as the ER and plasma membrane. We have solved the crystal structure of the ORP8 lipid transport domain (ORD8). The ORD8 exhibite...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10417380/ https://www.ncbi.nlm.nih.gov/pubmed/37566053 http://dx.doi.org/10.3390/cells12151974 |
_version_ | 1785088019352518656 |
---|---|
author | Eisenreichova, Andrea Klima, Martin Anila, Midhun Mohan Koukalova, Alena Humpolickova, Jana Różycki, Bartosz Boura, Evzen |
author_facet | Eisenreichova, Andrea Klima, Martin Anila, Midhun Mohan Koukalova, Alena Humpolickova, Jana Różycki, Bartosz Boura, Evzen |
author_sort | Eisenreichova, Andrea |
collection | PubMed |
description | ORPs are lipid-transport proteins belonging to the oxysterol-binding protein family. They facilitate the transfer of lipids between different intracellular membranes, such as the ER and plasma membrane. We have solved the crystal structure of the ORP8 lipid transport domain (ORD8). The ORD8 exhibited a β-barrel fold composed of anti-parallel β-strands, with three α-helices replacing β-strands on one side. This mixed alpha–beta structure was consistent with previously solved structures of ORP2 and ORP3. A large cavity (≈1860 Å(3)) within the barrel was identified as the lipid-binding site. Although we were not able to obtain a lipid-bound structure, we used computer simulations based on our crystal structure to dock PS and PI4P molecules into the putative lipid-binding site of the ORD8. Comparative experiments between the short ORD8(ΔLid) (used for crystallography) and the full-length ORD8 (lid containing) revealed the lid’s importance for stable lipid binding. Fluorescence assays revealed different transport efficiencies for PS and PI4P, with the lid slowing down transport and stabilizing cargo. Coarse-grained simulations highlighted surface-exposed regions and hydrophobic interactions facilitating lipid bilayer insertion. These findings enhance our comprehension of ORD8, its structure, and lipid transport mechanisms, as well as provide a structural basis for the design of potential inhibitors. |
format | Online Article Text |
id | pubmed-10417380 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-104173802023-08-12 Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport Eisenreichova, Andrea Klima, Martin Anila, Midhun Mohan Koukalova, Alena Humpolickova, Jana Różycki, Bartosz Boura, Evzen Cells Article ORPs are lipid-transport proteins belonging to the oxysterol-binding protein family. They facilitate the transfer of lipids between different intracellular membranes, such as the ER and plasma membrane. We have solved the crystal structure of the ORP8 lipid transport domain (ORD8). The ORD8 exhibited a β-barrel fold composed of anti-parallel β-strands, with three α-helices replacing β-strands on one side. This mixed alpha–beta structure was consistent with previously solved structures of ORP2 and ORP3. A large cavity (≈1860 Å(3)) within the barrel was identified as the lipid-binding site. Although we were not able to obtain a lipid-bound structure, we used computer simulations based on our crystal structure to dock PS and PI4P molecules into the putative lipid-binding site of the ORD8. Comparative experiments between the short ORD8(ΔLid) (used for crystallography) and the full-length ORD8 (lid containing) revealed the lid’s importance for stable lipid binding. Fluorescence assays revealed different transport efficiencies for PS and PI4P, with the lid slowing down transport and stabilizing cargo. Coarse-grained simulations highlighted surface-exposed regions and hydrophobic interactions facilitating lipid bilayer insertion. These findings enhance our comprehension of ORD8, its structure, and lipid transport mechanisms, as well as provide a structural basis for the design of potential inhibitors. MDPI 2023-07-31 /pmc/articles/PMC10417380/ /pubmed/37566053 http://dx.doi.org/10.3390/cells12151974 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Eisenreichova, Andrea Klima, Martin Anila, Midhun Mohan Koukalova, Alena Humpolickova, Jana Różycki, Bartosz Boura, Evzen Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title | Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title_full | Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title_fullStr | Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title_full_unstemmed | Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title_short | Crystal Structure of the ORP8 Lipid Transport ORD Domain: Model of Lipid Transport |
title_sort | crystal structure of the orp8 lipid transport ord domain: model of lipid transport |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10417380/ https://www.ncbi.nlm.nih.gov/pubmed/37566053 http://dx.doi.org/10.3390/cells12151974 |
work_keys_str_mv | AT eisenreichovaandrea crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT klimamartin crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT anilamidhunmohan crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT koukalovaalena crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT humpolickovajana crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT rozyckibartosz crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport AT bouraevzen crystalstructureoftheorp8lipidtransportorddomainmodeloflipidtransport |