2m7x
From Proteopedia
Structural and Functional Analysis of Transmembrane Segment IV of the Salt Tolerance Protein Sod2
Structural highlights
FunctionNAH_SCHPO Sodium export from cell, takes up external protons in exchange for internal sodium ions. Involved in regulation of pH. Publication Abstract from PubMedSod2 is the plasma membrane Na+/H+ exchanger of fission yeast S. pombe. It provides salt tolerance by removing excess intracellular sodium (or lithium) in exchange for protons. We examined the role of amino acid residues of transmembrane segment IV (TM IV) (126FPQINFLGSLLIAGCITSTDPVLSALI152) in activity by using alanine scanning mutagenesis and examining salt tolerance in sod2-deficient S. pombe. Two amino acids were critical for function. Mutations T144A and V147A resulted in defective proteins that did not confer salt tolerance when re-introduced into S. pombe. Sod2 protein with other alanine mutations in TM IV, had little or no effects. T144D and T144K mutant proteins were inactive, however a T144S protein was functional and provided lithium, but not sodium, tolerance and transport. Analysis of sensitivity to trypsin indicated that the mutations caused a conformational change in the sod2 protein. We expressed and purified TM IV (amino acids 125-154). NMR analysis yielded a model with two helical regions (amino acids 128-142 and 147-154) separated by unwound region (amino acids 143-146). Molecular modeling of the entire sod2 protein suggested TM IV has a structure similar to that deduced by NMR analysis and an overall structure similar to that of E. coli NhaA. TM IV of sod2 has similarities to TM V of the Zygosaccharomyces rouxii Na+/H+ exchanger and TM VI of isoform 1 of mammalian Na+/H+ exchanger. TM IV of sod2 is critical to transport and may be involved in cation binding or conformational changes of the protein. Structural and Functional Analysis of Transmembrane Segment IV of the Salt Tolerance Protein Sod2.,Ullah A, Kemp G, Lee B, Alves C, Young H, Sykes BD, Fliegel L J Biol Chem. 2013 Jul 8. PMID:23836910[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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