Substrate binding and reactivity are not linked: grafting a proton-transfer network into a Class 1A dihydroorotate dehydrogenase.
Substrate binding and reactivity are not linked: grafting a proton-transfer network into a Class 1A dihydroorotate dehydrogenase.
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底物结合和反应性不相关：将质子转移网络移植到 1A 类二氢乳清酸脱氢酶中。
DOI：
10.1021/bi200258y
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发表时间：
2011
期刊：
影响因子：
2.9
通讯作者：
Palfey,BruceA
中科院分区：
文献类型：
--
作者：
McDonald,ClaudiaA;Palfey,BruceA
添加两个残基，包括保守的质子转移网络的2类二氢乳清酸脱氢酶（DHOD）的Cys130Ser类1A DHOD没有恢复的功能的活性位点基地或快速黄素还原。对三重、双重和单突变1A类酶的研究表明，网络实际上阻止了半胱氨酸作为碱基，并且网络残基独立地起作用。我们的数据表明，残基71是配体结合特异性的重要决定因素。Leu71Phe突变收紧二氢乳清酸结合，但削弱1A类酶的苯甲酸盐抑制剂的结合。
Adding the two residues comprising the conserved proton-transfer network of Class 2 dihydroorotate dehydrogenase (DHOD) to the Cys130Ser Class 1A DHOD did not restore the function of the active site base or rapid flavin reduction. Studies of triple, double, and single mutant Class 1A enzymes showed that the network actually prevents cysteine from acting as a base and that the network residues act independently. Our data show that residue 71 is an important determinant of ligand binding specificity. The Leu71Phe mutation tightens dihydrooroate binding but weakens the binding of benzoate inhibitors of Class 1A enzymes.