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Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase
P-type ATPases of subfamily IV (P4-ATPases) constitute a major group of phospholipid flippases that form heteromeric complexes with members of the Cdc50 (cell division control 50) protein family. Some P4-ATPases interact specifically with only one β-subunit isoform, whereas others are promiscuous an...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Portland Press Ltd.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888458/ https://www.ncbi.nlm.nih.gov/pubmed/27048590 http://dx.doi.org/10.1042/BCJ20160207 |
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author | Costa, Sara R. Marek, Magdalena Axelsen, Kristian B. Theorin, Lisa Pomorski, Thomas G. López-Marqués, Rosa L. |
author_facet | Costa, Sara R. Marek, Magdalena Axelsen, Kristian B. Theorin, Lisa Pomorski, Thomas G. López-Marqués, Rosa L. |
author_sort | Costa, Sara R. |
collection | PubMed |
description | P-type ATPases of subfamily IV (P4-ATPases) constitute a major group of phospholipid flippases that form heteromeric complexes with members of the Cdc50 (cell division control 50) protein family. Some P4-ATPases interact specifically with only one β-subunit isoform, whereas others are promiscuous and can interact with several isoforms. In the present study, we used a site-directed mutagenesis approach to assess the role of post-translational modifications at the plant ALIS5 β-subunit ectodomain in the functionality of the promiscuous plant P4-ATPase ALA2. We identified two N-glycosylated residues, Asn(181) and Asn(231). Whereas mutation of Asn(231) seems to have a small effect on P4-ATPase complex formation, mutation of evolutionarily conserved Asn(181) disrupts interaction between the two subunits. Of the four cysteine residues located in the ALIS5 ectodomain, mutation of Cys(86) and Cys(107) compromises complex association, but the mutant β-subunits still promote complex trafficking and activity to some extent. In contrast, disruption of a conserved disulfide bond between Cys(158) and Cys(172) has no effect on the P4-ATPase complex. Our results demonstrate that post-translational modifications in the β-subunit have different functional roles in different organisms, which may be related to the promiscuity of the P4-ATPase. |
format | Online Article Text |
id | pubmed-4888458 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Portland Press Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-48884582016-06-08 Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase Costa, Sara R. Marek, Magdalena Axelsen, Kristian B. Theorin, Lisa Pomorski, Thomas G. López-Marqués, Rosa L. Biochem J Research Articles P-type ATPases of subfamily IV (P4-ATPases) constitute a major group of phospholipid flippases that form heteromeric complexes with members of the Cdc50 (cell division control 50) protein family. Some P4-ATPases interact specifically with only one β-subunit isoform, whereas others are promiscuous and can interact with several isoforms. In the present study, we used a site-directed mutagenesis approach to assess the role of post-translational modifications at the plant ALIS5 β-subunit ectodomain in the functionality of the promiscuous plant P4-ATPase ALA2. We identified two N-glycosylated residues, Asn(181) and Asn(231). Whereas mutation of Asn(231) seems to have a small effect on P4-ATPase complex formation, mutation of evolutionarily conserved Asn(181) disrupts interaction between the two subunits. Of the four cysteine residues located in the ALIS5 ectodomain, mutation of Cys(86) and Cys(107) compromises complex association, but the mutant β-subunits still promote complex trafficking and activity to some extent. In contrast, disruption of a conserved disulfide bond between Cys(158) and Cys(172) has no effect on the P4-ATPase complex. Our results demonstrate that post-translational modifications in the β-subunit have different functional roles in different organisms, which may be related to the promiscuity of the P4-ATPase. Portland Press Ltd. 2016-05-27 2016-06-01 /pmc/articles/PMC4888458/ /pubmed/27048590 http://dx.doi.org/10.1042/BCJ20160207 Text en © 2016 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society |
spellingShingle | Research Articles Costa, Sara R. Marek, Magdalena Axelsen, Kristian B. Theorin, Lisa Pomorski, Thomas G. López-Marqués, Rosa L. Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title | Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title_full | Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title_fullStr | Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title_full_unstemmed | Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title_short | Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase |
title_sort | role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant p4-atpase |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888458/ https://www.ncbi.nlm.nih.gov/pubmed/27048590 http://dx.doi.org/10.1042/BCJ20160207 |
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