Arg901 in the AE1 C-terminal tail is involved in conformational change but not in substrate binding
Arg901 in the AE1 C-terminal tail is involved in conformational change but not in substrate binding
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AE1 C 末端尾部的 Arg901 参与构象变化，但不参与底物结合
DOI：
10.1016/j.bbamem.2011.11.019
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Hamasaki N
中科院分区：
文献类型：
--
作者：
Takazaki;S;Abe;Y;Yamaguchi T;Yagi M;Ueda T;Kagn D;Hamasaki N
在我们之前的论文中，我们证明了人AE 1（带3，阴离子交换剂1）C-末端尾部的Arg 901在阴离子交换过程中的构象变化中具有功能作用。为了进一步研究精氨酸901是如何参与构象变化的，我们在酿酒酵母的质膜上表达了各种精氨酸901突变体和C-末端尾的丙氨酸突变体（从Leu 886到瓦尔911），并评估了硫酸根离子转运的动力学参数。因此，Vmax随着第901和外围亲水残基的疏水性增加而降低，表明C-末端残基的疏水性参与构象变化。我们还发现，Arg 901修饰后的C-末端区域的碱性和蛋白酶的阻力与羟基苯乙二醛（HPG）或苯乙二醛（PG），疏水试剂。这些结果表明，精氨酸901周围的C-末端区域的疏水性增加导致新产生的疏水相互作用导致低效的构象变化。
In our previous paper, we demonstrated that Arg 901 in the C-terminal tail of human AE1 (band 3, anion exchanger 1) had a functional role in conformational change during anion exchange. To further examine how Arg 901 is involved in conformational change, we expressed various Arg 901 mutants and alanine mutants of the C-terminal tail (from Leu 886 to Val 911) on the plasma membrane of Saccharomyces cerevisiae and evaluated the kinetic parameters of sulfate ion transport. As a result, Vmax decreased as the hydrophobicities of the 901st and peripheral hydrophilic residues increased, indicating that the hydrophobicity of the C-terminal residue is involved in the conformational change. We also found the alkali and protease resistance of the C-terminal region after Arg 901 modification with hydroxyphenylglyoxal (HPG) or phenylglyoxal (PG), a hydrophobic reagent. These results suggested that the increased hydrophobicity of the C-terminal region around Arg 901 leads to inefficient conformational change by the newly produced hydrophobic interaction.