Molecular and functional characterization of grapevine NIPs through heterologous expression in aqy-null Saccharomyces cerevisiae
| Autor: | Farzana Sabir, Catarina Prista, Sara C. Gomes, Maria C. Loureiro-Dias, Graça Soveral |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine metalloids nips Saccharomyces cerevisiae Glycerol transport Aquaporin glycerol 01 natural sciences Catalysis Inorganic Chemistry saccharomyces cerevisiae lcsh:Chemistry 03 medical and health sciences chemistry.chemical_compound NIPs Glycerol Physical and Theoretical Chemistry Molecular Biology lcsh:QH301-705.5 Spectroscopy biology Chemistry Organic Chemistry General Medicine biology.organism_classification Yeast 3. Good health Computer Science Applications grapevine aquaporin Cytosol 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 Biophysics Membrane channel Heterologous expression 010606 plant biology & botany |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP International Journal of Molecular Sciences, Vol 21, Iss 2, p 663 (2020) International Journal of Molecular Sciences Volume 21 Issue 2 |
| Popis: | Plant Nodulin 26-like Intrinsic Proteins (NIPs) are multifunctional membrane channels of the Major Intrinsic Protein (MIP) family. Unlike other homologs, they have low intrinsic water permeability. NIPs possess diverse substrate selectivity, ranging from water to glycerol and to other small solutes, depending on the group-specific amino acid composition at aromatic/Arg (ar/R) constriction. We cloned three NIPs (NIP1 1, NIP5 1, and NIP6 1) from grapevine (cv. Touriga Nacional). Their expression in the membrane of aqy-null Saccharomyces cerevisiae enabled their functional characterization for water and glycerol transport through stopped-flow spectroscopy. VvTnNIP1 1 demonstrated high water as well as glycerol permeability, whereas VvTnNIP6 1 was impermeable to water but presented high glycerol permeability. Their transport activities were declined by cytosolic acidification, implying that internal-pH can regulate NIPs gating. Furthermore, an extension of C-terminal in VvTnNIP6 1M homolog, led to improved channel activity, suggesting that NIPs gating is putatively regulated by C-terminal. Yeast growth assays in the presence of diverse substrates suggest that the transmembrane flux of metalloids (As, B, and Se) and the heavy metal (Cd) are facilitated through grapevine NIPs. This is the first molecular and functional characterization of grapevine NIPs, providing crucial insights into understanding their role for uptake and translocation of small solutes, and extrusion of toxic compounds in grapevine. |
| Databáze: | OpenAIRE |
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