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https://hdl.handle.net/2440/70399
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Type: | Journal article |
Title: | Structural and functional analyses of PpENA1 provide insights into cation binding by type IID P-type ATPases in lower plants and fungi |
Author: | Drew, D. Hrmova, M. Lunde, C. Jacobs, A. Tester, M. Fincher, G. |
Citation: | BBA: Biomembranes, 2011; 1808(6):1483-1492 |
Publisher: | Elsevier Science BV |
Issue Date: | 2011 |
ISSN: | 0005-2736 1879-2642 |
Statement of Responsibility: | Damian P. Drew, Maria Hrmova, Christina Lunde, Andrew K. Jacobs, Mark Tester and Geoffrey B. Fincher |
Abstract: | PpENA1 is a membrane-spanning transporter from the moss Physcomitrella patens, and is the first type IID P-type ATPase to be reported in the plant kingdom. In Physcomitrella, PpENA1 is essential for normal growth under moderate salt stress, while in yeast, type IID ATPases provide a vital efflux mechanism for cells under high salt conditions by selectively transporting Na+ or K+ across the plasma membrane. To investigate the structural basis for cation-binding within the type IID ATPase subfamily, we used homology modeling to identify a highly conserved cation-binding pocket between membrane helix (MH) 4 and MH 6 of the membrane-spanning pore of PpENA1. Mutation of specific charged and polar residues on MHs 4-6 resulted in a decrease or loss of protein activity as measured by complementation assays in yeast. The E298S mutation on MH 4 of PpENA1 had the most significant effect on activity despite the presence of a serine at this position in fungal type IID ATPases. Activity was partially restored in an inactivated PpENA1 mutant by the insertion of two additional serine residues on MH 4 and one on MH 6 based on the presence of these residues in fungal type IID ATPases. Our results suggest that the residues responsible for cation-binding in PpENA1 are distinct from those in fungal type IID ATPases, and that a fungal-type cation binding site can be successfully engineered into the moss protein. |
Keywords: | Fungi Saccharomyces cerevisiae Bryopsida Cations Potassium Sodium Fungal Proteins Plant Proteins Immunoblotting Genetic Complementation Test Amino Acid Substitution Binding Sites Ion Transport Amino Acid Sequence Protein Structure, Secondary Protein Structure, Tertiary Protein Binding Sequence Homology, Amino Acid Mutation Models, Molecular Molecular Sequence Data Sodium-Potassium-Exchanging ATPase |
Rights: | © 2010 Elsevier B.V. All rights reserved. |
DOI: | 10.1016/j.bbamem.2010.11.013 |
Grant ID: | ARC |
Published version: | http://dx.doi.org/10.1016/j.bbamem.2010.11.013 |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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