Identification of Conserved Polar Residues Important for Salt Tolerance by the Na+/H+ Exchanger of Schizosaccharomyces Pombe

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 2005 Feb 24

PMID 15724441

Citations 4

Authors

Larry Fliegel

Affiliations

Soon will be listed here.

Abstract

The Na+/H+ exchanger is a ubiquitous protein that transports Na+ and H+ in opposite directions across cell membranes. In fission yeast, the Na+/H+ exchanger sod2 plays a major role in the removal of excess detrimental intracellular sodium. The effect of mutagenesis of conserved polar amino acids of sod2 was examined by expressing 10 different mutant forms of sod2 in sod2 deficient S. pombe and characterizing salt tolerance. Asp145, 266, 267, and Glu173 were critical for proper function of sod2. Asp241 had an intermediate effect on sod2 function while mutation of Asp178 did not impair sod2 function. Simultaneous mutation of the Asp266, 267 pair impaired sod2 function. Mutation of each individual residue demonstrated that both were critical for sod2 function. Conservative mutations (Asp to Glu) of Asp266 and 267 failed to restore sod2 function. The results suggest that acidic residues associated with transmembrane segments are important in function, possibly being important in binding and coordinating cations.

Citing Articles

Expression and characterization of the SOS1 Arabidopsis salt tolerance protein.

Ullah A, Dutta D, Fliegel L Mol Cell Biochem. 2016; 415(1-2):133-43.

PMID: 26992907 DOI: 10.1007/s11010-016-2685-2.

Functional role and analysis of cysteine residues of the salt tolerance protein Sod2.

Ullah A, El-Magd R, Fliegel L Mol Cell Biochem. 2013; 386(1-2):85-98.

PMID: 24104454 DOI: 10.1007/s11010-013-1847-8.

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 B J Biol Chem. 2013; 288(34):24609-24.

PMID: 23836910 PMC: 3750159. DOI: 10.1074/jbc.M113.483065.

Alkali metal cation transport and homeostasis in yeasts.

Arino J, Ramos J, Sychrova H Microbiol Mol Biol Rev. 2010; 74(1):95-120.

PMID: 20197501 PMC: 2832347. DOI: 10.1128/MMBR.00042-09.

References

Lee J, Varela M, Wilson T . Physiological evidence for an interaction between Glu-325 and His-322 in the lactose carrier of Escherichia coli. Biochim Biophys Acta. 1996; 1278(1):111-8. DOI: 10.1016/0005-2736(95)00209-x. View

Nakamura T, Komano Y, Itaya E, Tsukamoto K, Tsuchiya T, UNEMOTO T . Cloning and sequencing of an Na+/H+ antiporter gene from the marine bacterium Vibrio alginolyticus. Biochim Biophys Acta. 1994; 1190(2):465-8. DOI: 10.1016/0005-2736(94)90109-0. View

Sardet C, Franchi A, Pouyssegur J . Molecular cloning, primary structure, and expression of the human growth factor-activatable Na+/H+ antiporter. Cell. 1989; 56(2):271-80. DOI: 10.1016/0092-8674(89)90901-x. View

Mogi T, Stern L, Marti T, Chao B, Khorana H . Aspartic acid substitutions affect proton translocation by bacteriorhodopsin. Proc Natl Acad Sci U S A. 1988; 85(12):4148-52. PMC: 280383. DOI: 10.1073/pnas.85.12.4148. View

Galili L, Rothman A, Kozachkov L, Rimon A, Padan E . Trans membrane domain IV is involved in ion transport activity and pH regulation of the NhaA-Na(+)/H(+) antiporter of Escherichia coli. Biochemistry. 2002; 41(2):609-17. DOI: 10.1021/bi011655v. View

Padan E, Venturi M, Gerchman Y, Dover N . Na(+)/H(+) antiporters. Biochim Biophys Acta. 2001; 1505(1):144-57. DOI: 10.1016/s0005-2728(00)00284-x. View

Dibrov P, Smith J, Young P, Fliegel L . Identification and localization of the sod2 gene product in fission yeast. FEBS Lett. 1997; 405(1):119-24. DOI: 10.1016/s0014-5793(97)00169-5. View

Nass R, Rao R . Novel localization of a Na+/H+ exchanger in a late endosomal compartment of yeast. Implications for vacuole biogenesis. J Biol Chem. 1998; 273(33):21054-60. DOI: 10.1074/jbc.273.33.21054. View

Kuroda T, Shimamoto T, Inaba K, Tsuda M, Tsuchiya T . Properties and sequence of the NhaA Na+/H+ antiporter of Vibrio parahaemolyticus. J Biochem. 1994; 116(5):1030-8. DOI: 10.1093/oxfordjournals.jbchem.a124624. View

10.

Nakamura T, Komano Y, UNEMOTO T . Three aspartic residues in membrane-spanning regions of Na+/H+ antiporter from Vibrio alginolyticus play a role in the activity of the carrier. Biochim Biophys Acta. 1995; 1230(3):170-6. DOI: 10.1016/0005-2728(95)00053-l. View

11.

Otto H, Marti T, Holz M, Mogi T, Lindau M, Khorana H . Aspartic acid-96 is the internal proton donor in the reprotonation of the Schiff base of bacteriorhodopsin. Proc Natl Acad Sci U S A. 1989; 86(23):9228-32. PMC: 298467. DOI: 10.1073/pnas.86.23.9228. View

12.

Inoue H, Noumi T, Tsuchiya T, Kanazawa H . Essential aspartic acid residues, Asp-133, Asp-163 and Asp-164, in the transmembrane helices of a Na+/H+ antiporter (NhaA) from Escherichia coli. FEBS Lett. 1995; 363(3):264-8. DOI: 10.1016/0014-5793(95)00331-3. View

13.

Wang Z, Orlowski J, Shull G . Primary structure and functional expression of a novel gastrointestinal isoform of the rat Na/H exchanger. J Biol Chem. 1993; 268(16):11925-8. View

14.

Watanabe Y, Shimono Y, Tsuji H, Tamai Y . Role of the glutamic and aspartic residues in Na+-ATPase function in the ZrENA1 gene of Zygosaccharomyces rouxii. FEMS Microbiol Lett. 2002; 209(1):39-43. DOI: 10.1111/j.1574-6968.2002.tb11106.x. View

15.

Watanabe Y, Miwa S, Tamai Y . Characterization of Na+/H(+)-antiporter gene closely related to the salt-tolerance of yeast Zygosaccharomyces rouxii. Yeast. 1995; 11(9):829-38. DOI: 10.1002/yea.320110905. View

16.

Dibrov P, Young P, Fliegel L . Functional analysis of amino acid residues essential for activity in the Na+/H+ exchanger of fission yeast. Biochemistry. 1998; 37(23):8282-8. DOI: 10.1021/bi9801457. View

17.

Jia Z, McCullough N, Martel R, Hemmingsen S, Young P . Gene amplification at a locus encoding a putative Na+/H+ antiporter confers sodium and lithium tolerance in fission yeast. EMBO J. 1992; 11(4):1631-40. PMC: 556613. DOI: 10.1002/j.1460-2075.1992.tb05209.x. View

18.

Nicoll D, Hryshko L, Matsuoka S, Frank J, Philipson K . Mutation of amino acid residues in the putative transmembrane segments of the cardiac sarcolemmal Na+-Ca2+ exchanger. J Biol Chem. 1996; 271(23):13385-91. DOI: 10.1074/jbc.271.23.13385. View

19.

Murtazina R, Booth B, Bullis B, Singh D, Fliegel L . Functional analysis of polar amino-acid residues in membrane associated regions of the NHE1 isoform of the mammalian Na+/H+ exchanger. Eur J Biochem. 2001; 268(17):4674-85. DOI: 10.1046/j.1432-1327.2001.02391.x. View

20.

Wakabayashi S, Pang T, Su X, Shigekawa M . A novel topology model of the human Na(+)/H(+) exchanger isoform 1. J Biol Chem. 2000; 275(11):7942-9. DOI: 10.1074/jbc.275.11.7942. View