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首页> 外文期刊>Biochemistry >Functional analysis of amino acid residues essential for activity in the Na+/H+ exchanger of fission yeast
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Functional analysis of amino acid residues essential for activity in the Na+/H+ exchanger of fission yeast

机译：裂变酵母Na + / H +交换子中活性必不可少的氨基酸残基的功能分析

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We identified amino acid residues important for activity of sod2, the Na+/H+ antiporter of Schizosaccharomyces pombe. We mutated all eight His residues of sod2 into Arg. Only His367-->Arg affected function and resulted in complete inability of sod2 to allow growth of S. pombe in LiCl-containing medium. Mutant S. pombe (H367R) could not expel sodium in acidic (pH 4.0) medium and were defective in their ability to alkalinize external medium. When His367 was replaced by Asp, sodium export of S. pombe was suppressed at acidic pH while the sodium-dependent proton influx at pH 6.1 was increased compared to wild type. We also mutated three residues conserved in putative membrane regions of various eukaryotic and prokaryotic Na+/H+ exchangers. S. pombe containing Asp241-->Asn and Asp266,267-->Asn mutations had greatly impaired growth in LiCl-containing medium. In addition, sodium-dependent proton influx at external pH 6.1 was Impaired. Sodium export from S. pombe cells at external pH 4.0 was also almost completely abolished by the D266,267N mutation; however, the D241N mutant protein retained almost normal Na+ export. The results demonstrate that His367, Asp241, and Asp266,267 are important in the function of the eukaryotic Na+/H+ exchanger sod2. [References: 29]
机译：我们鉴定了对粟酒裂殖酵母的Na + / H +反转运蛋白sod2活性重要的氨基酸残基。我们将sod2的所有八个His残基突变为Arg。只有His367-> Arg会影响功能，并导致sod2完全无法使粟酒裂殖酵母在含LiCl的培养基中生长。突变体粟酒裂殖酵母（H367R）不能在酸性（pH 4.0）培养基中排出钠盐，并且不能碱化外部培养基。与野生型相比，当His367替换为Asp时，粟酒裂殖酵母的钠出口在酸性pH受到抑制，而在pH 6.1的钠依赖性质子流入增加。我们还突变了各种真核和原核Na + / H +交换子的推定膜区域中保守的三个残基。含有Asp241-> Asn和Asp266,267-> Asn突变的粟酒裂殖酵母大大削弱了含LiCl培养基的生长。此外，在外部pH 6.1时钠依赖性质子涌入受到损害。 D266,267N突变也几乎完全消除了外部pH 4.0下从粟酒裂殖酵母细胞输出的钠。然而，D241N突变蛋白保留了几乎正常的Na +输出。结果表明，His367，Asp241和Asp266,267在真核Na + / H +交换子sod2的功能中很重要。 [参考：29]

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