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首页> 外文期刊>Biochemistry >Role of individual positive charges in the membrane orientation and activity of transporters of the small multidrug resistance family
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Role of individual positive charges in the membrane orientation and activity of transporters of the small multidrug resistance family

机译：单个正电荷在小型多药耐药家族的膜取向和转运蛋白活性中的作用

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摘要

The effect of individual positively charged residues on the orientation in the membrane was analyzed in three dual-topology transporters of the small multidrug resistance (SMR) family: AAVE4701aave of Acidovorax avenae, EMREecol of Escherichia coli, and RRUA0272rrub of Rhodospirillum rubrum. It is shown that (i) individual positive charges have different impacts on the orientation, (ii) positive charges that are conserved in the three different proteins do not have the same impact on the orientation, (iii) positive charges in odd- and even-numbered loops have different impacts, (iv) for some, but not all, the impact depends on the presence of other positive charges, and (v) proteins from which all positive charges are removed in some cases are dual-topology proteins and in other cases have a single orientation. A small number of positive charges placed in the loops of the latter proteins results in the violation of the so-called positive-inside rule that has been reported previously [Kolbusz, M. A., et al. (2010) J. Mol. Biol. 402, 127-138]. We conclude that each positive charge shifts the distribution between the two orientations toward the state that has the positive charge in the cytoplasm but that intrinsic factors other than positive charges determine the orientation as well. The ability of the mutants of AAVE4701aave and EMREecol to confer resistance against ethidium bromide revealed an essential role in catalysis for a conserved pair of positive charges in the second loop. No significant relation between activity and the relative orientation of the monomeric subunits in the dimer could be demonstrated.
机译：在小型多药耐药性（SMR）家族的三个双拓扑转运蛋白中分析了单个带正电荷的残基对膜取向的影响：酸性菌AAVE4701aave，大肠杆菌EMREecol和红斑霉菌RRUA0272rrub。结果表明：（i）单个正电荷对方向的影响不同;（ii）三种不同蛋白质中保守的正电荷对方向的影响不同;（iii）奇数和偶数的正电荷编号的环具有不同的影响，（iv）对某些（但不是全部）影响取决于其他正电荷的存在，并且（v）在某些情况下从中除去所有正电荷的蛋白是双拓扑蛋白，并且其他情况有一个单一的方向。置于后者蛋白质环中的少量正电荷会违反先前报道的所谓的正内规则[Kolbusz，M. A.，et al。 （2010）J.Mol。生物学402，127-138]。我们得出的结论是，每个正电荷都将两个方向之间的分布向着在细胞质中具有正电荷的状态转移，但除正电荷以外的其他内在因素也决定了方向。 AAVE4701aave和EMREecol突变体赋予对溴化乙锭抗性的能力揭示了在第二环中一对保守的正电荷催化中的重要作用。活性与二聚体中单体亚基的相对取向之间没有显着关系。

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