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Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins
In Arabidopsis, vacuolar sorting receptor isoform 1 (VSR1) sorts 12S globulins to the protein storage vacuoles during seed development. Vacuolar sorting is mediated by specific protein–protein interactions between VSR1 and the vacuolar sorting determinant located at the C terminus (ctVSD) on the car...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8740768/ https://www.ncbi.nlm.nih.gov/pubmed/34983843 http://dx.doi.org/10.1073/pnas.2111281119 |
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author | Tsao, Hsi-En Lui, Shu Nga Lo, Anthony Hiu-Fung Chen, Shuai Wong, Hiu Yan Wong, Chi-Kin Jiang, Liwen Wong, Kam-Bo |
author_facet | Tsao, Hsi-En Lui, Shu Nga Lo, Anthony Hiu-Fung Chen, Shuai Wong, Hiu Yan Wong, Chi-Kin Jiang, Liwen Wong, Kam-Bo |
author_sort | Tsao, Hsi-En |
collection | PubMed |
description | In Arabidopsis, vacuolar sorting receptor isoform 1 (VSR1) sorts 12S globulins to the protein storage vacuoles during seed development. Vacuolar sorting is mediated by specific protein–protein interactions between VSR1 and the vacuolar sorting determinant located at the C terminus (ctVSD) on the cargo proteins. Here, we determined the crystal structure of the protease-associated domain of VSR1 (VSR1-PA) in complex with the C-terminal pentapeptide ((468)RVAAA(472)) of cruciferin 1, an isoform of 12S globulins. The (468)RVA(470) motif forms a parallel β-sheet with the switch III residues ((127)TMD(129)) of VSR1-PA, and the (471)AA(472) motif docks to a cradle formed by the cargo-binding loop ((95)RGDCYF(100)), making a hydrophobic interaction with Tyr99. The C-terminal carboxyl group of the ctVSD is recognized by forming salt bridges with Arg95. The C-terminal sequences of cruciferin 1 and vicilin-like storage protein 22 were sufficient to redirect the secretory red fluorescent protein (spRFP) to the vacuoles in Arabidopsis protoplasts. Adding a proline residue to the C terminus of the ctVSD and R95M substitution of VSR1 disrupted receptor–cargo interactions in vitro and led to increased secretion of spRFP in Arabidopsis protoplasts. How VSR1-PA recognizes ctVSDs of other storage proteins was modeled. The last three residues of ctVSD prefer hydrophobic residues because they form a hydrophobic cluster with Tyr99 of VSR1-PA. Due to charge–charge interactions, conserved acidic residues, Asp129 and Glu132, around the cargo-binding site should prefer basic residues over acidic ones in the ctVSD. The structural insights gained may be useful in targeting recombinant proteins to the protein storage vacuoles in seeds. |
format | Online Article Text |
id | pubmed-8740768 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-87407682022-06-30 Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins Tsao, Hsi-En Lui, Shu Nga Lo, Anthony Hiu-Fung Chen, Shuai Wong, Hiu Yan Wong, Chi-Kin Jiang, Liwen Wong, Kam-Bo Proc Natl Acad Sci U S A Biological Sciences In Arabidopsis, vacuolar sorting receptor isoform 1 (VSR1) sorts 12S globulins to the protein storage vacuoles during seed development. Vacuolar sorting is mediated by specific protein–protein interactions between VSR1 and the vacuolar sorting determinant located at the C terminus (ctVSD) on the cargo proteins. Here, we determined the crystal structure of the protease-associated domain of VSR1 (VSR1-PA) in complex with the C-terminal pentapeptide ((468)RVAAA(472)) of cruciferin 1, an isoform of 12S globulins. The (468)RVA(470) motif forms a parallel β-sheet with the switch III residues ((127)TMD(129)) of VSR1-PA, and the (471)AA(472) motif docks to a cradle formed by the cargo-binding loop ((95)RGDCYF(100)), making a hydrophobic interaction with Tyr99. The C-terminal carboxyl group of the ctVSD is recognized by forming salt bridges with Arg95. The C-terminal sequences of cruciferin 1 and vicilin-like storage protein 22 were sufficient to redirect the secretory red fluorescent protein (spRFP) to the vacuoles in Arabidopsis protoplasts. Adding a proline residue to the C terminus of the ctVSD and R95M substitution of VSR1 disrupted receptor–cargo interactions in vitro and led to increased secretion of spRFP in Arabidopsis protoplasts. How VSR1-PA recognizes ctVSDs of other storage proteins was modeled. The last three residues of ctVSD prefer hydrophobic residues because they form a hydrophobic cluster with Tyr99 of VSR1-PA. Due to charge–charge interactions, conserved acidic residues, Asp129 and Glu132, around the cargo-binding site should prefer basic residues over acidic ones in the ctVSD. The structural insights gained may be useful in targeting recombinant proteins to the protein storage vacuoles in seeds. National Academy of Sciences 2021-12-30 2022-01-04 /pmc/articles/PMC8740768/ /pubmed/34983843 http://dx.doi.org/10.1073/pnas.2111281119 Text en Copyright © 2021 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences Tsao, Hsi-En Lui, Shu Nga Lo, Anthony Hiu-Fung Chen, Shuai Wong, Hiu Yan Wong, Chi-Kin Jiang, Liwen Wong, Kam-Bo Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title | Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title_full | Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title_fullStr | Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title_full_unstemmed | Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title_short | Structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
title_sort | structural insights into how vacuolar sorting receptors recognize the sorting determinants of seed storage proteins |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8740768/ https://www.ncbi.nlm.nih.gov/pubmed/34983843 http://dx.doi.org/10.1073/pnas.2111281119 |
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