Cargando…

Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization

Sucrose-phosphate phosphatase (SPP) catalyses the final step in the sucrose biosynthesis pathway. Arabidopsis thaliana genome codifies four SPP isoforms. In this study, the four Arabidopsis thaliana genes coding for SPP isoforms have been cloned, expressed in Escherichia coli and the kinetic and reg...

Descripción completa

Detalles Bibliográficos
Autores principales: Albi, Tomás, Ruiz, M. Teresa, de los Reyes, Pedro, Valverde, Federico, Romero, José M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5113954/
https://www.ncbi.nlm.nih.gov/pubmed/27855180
http://dx.doi.org/10.1371/journal.pone.0166308
_version_ 1782468268838092800
author Albi, Tomás
Ruiz, M. Teresa
de los Reyes, Pedro
Valverde, Federico
Romero, José M.
author_facet Albi, Tomás
Ruiz, M. Teresa
de los Reyes, Pedro
Valverde, Federico
Romero, José M.
author_sort Albi, Tomás
collection PubMed
description Sucrose-phosphate phosphatase (SPP) catalyses the final step in the sucrose biosynthesis pathway. Arabidopsis thaliana genome codifies four SPP isoforms. In this study, the four Arabidopsis thaliana genes coding for SPP isoforms have been cloned, expressed in Escherichia coli and the kinetic and regulatory properties of the purified enzymes analysed. SPP2 is the isoform showing the highest activity, with SPP3b and SPP3a showing lower activity levels. No activity was detected for SPP1. We propose that this lack of activity is probably due to the absence of an essential amino acid participating in catalysis and/or in the binding of the substrate, sucrose-6-phosphate (Suc6P). The expression patterns of Arabidopsis SPP genes indicate that SPP2 and SPP3b are the main isoforms expressed in different tissues and organs, although the non-catalytic SPP1 is the main isoform expressed in roots. Thus, SPP1 could have acquired new unknown functions. We also show that the three catalytically active SPPs from Arabidopsis are dimers. By generating a chimeric SPP composed of the monomeric cyanobacterial SPP fused to the higher plant non-catalytic S6PPc domain (from SPP2), we show that the S6PPc domain is responsible for SPP dimerization. This is the first experimental study on the functionality and gene expression pattern of all the SPPs from a single plant species.
format Online
Article
Text
id pubmed-5113954
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-51139542016-12-08 Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization Albi, Tomás Ruiz, M. Teresa de los Reyes, Pedro Valverde, Federico Romero, José M. PLoS One Research Article Sucrose-phosphate phosphatase (SPP) catalyses the final step in the sucrose biosynthesis pathway. Arabidopsis thaliana genome codifies four SPP isoforms. In this study, the four Arabidopsis thaliana genes coding for SPP isoforms have been cloned, expressed in Escherichia coli and the kinetic and regulatory properties of the purified enzymes analysed. SPP2 is the isoform showing the highest activity, with SPP3b and SPP3a showing lower activity levels. No activity was detected for SPP1. We propose that this lack of activity is probably due to the absence of an essential amino acid participating in catalysis and/or in the binding of the substrate, sucrose-6-phosphate (Suc6P). The expression patterns of Arabidopsis SPP genes indicate that SPP2 and SPP3b are the main isoforms expressed in different tissues and organs, although the non-catalytic SPP1 is the main isoform expressed in roots. Thus, SPP1 could have acquired new unknown functions. We also show that the three catalytically active SPPs from Arabidopsis are dimers. By generating a chimeric SPP composed of the monomeric cyanobacterial SPP fused to the higher plant non-catalytic S6PPc domain (from SPP2), we show that the S6PPc domain is responsible for SPP dimerization. This is the first experimental study on the functionality and gene expression pattern of all the SPPs from a single plant species. Public Library of Science 2016-11-17 /pmc/articles/PMC5113954/ /pubmed/27855180 http://dx.doi.org/10.1371/journal.pone.0166308 Text en © 2016 Albi et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Albi, Tomás
Ruiz, M. Teresa
de los Reyes, Pedro
Valverde, Federico
Romero, José M.
Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title_full Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title_fullStr Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title_full_unstemmed Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title_short Characterization of the Sucrose Phosphate Phosphatase (SPP) Isoforms from Arabidopsis thaliana and Role of the S6PPc Domain in Dimerization
title_sort characterization of the sucrose phosphate phosphatase (spp) isoforms from arabidopsis thaliana and role of the s6ppc domain in dimerization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5113954/
https://www.ncbi.nlm.nih.gov/pubmed/27855180
http://dx.doi.org/10.1371/journal.pone.0166308
work_keys_str_mv AT albitomas characterizationofthesucrosephosphatephosphatasesppisoformsfromarabidopsisthalianaandroleofthes6ppcdomainindimerization
AT ruizmteresa characterizationofthesucrosephosphatephosphatasesppisoformsfromarabidopsisthalianaandroleofthes6ppcdomainindimerization
AT delosreyespedro characterizationofthesucrosephosphatephosphatasesppisoformsfromarabidopsisthalianaandroleofthes6ppcdomainindimerization
AT valverdefederico characterizationofthesucrosephosphatephosphatasesppisoformsfromarabidopsisthalianaandroleofthes6ppcdomainindimerization
AT romerojosem characterizationofthesucrosephosphatephosphatasesppisoformsfromarabidopsisthalianaandroleofthes6ppcdomainindimerization