School of Life Sciences, University of Technology, Sydney, NSW 2007, Australia.
Int J Mol Sci. 2023 Jan 30;24(3):2624. doi: 10.3390/ijms24032624.
ATP-binding cassette (ABC) transporters comprise a large superfamily of primary active transporters, which are integral membrane proteins that couple energy to the uphill vectorial transport of substrates across cellular membranes, with concomitant hydrolysis of ATP. ABC transporters are found in all living organisms, coordinating mostly import in prokaryotes and export in eukaryotes. Unlike the highly conserved nucleotide binding domains (NBDs), sequence conservation in the transmembrane domains (TMDs) is low, with their divergent nature likely reflecting a need to accommodate a wide range of substrate types in terms of mass and polarity. An explosion in high resolution structural analysis over the past decade and a half has produced a wealth of structural information for ABCs. Based on the structures, a general mechanism for ABC transporters has been proposed, known as the Switch or Alternating Access Model, which holds that the NBDs are widely separated, with the TMDs and NBDs together forming an intracellular-facing inverted "V" shape. Binding of two ATPs and the substrate to the inward-facing conformation induces a transition to an outward conformation. Despite this apparent progress, certainty around the transport mechanism for any given ABC remains elusive. How substrate binding and transport is coupled to ATP binding and hydrolysis is not known, and there is a large body of biochemical and biophysical data that is at odds with the widely separated NBDs being a functional physiological state. An alternative Constant Contact model has been proposed in which the two NBSs operate 180 degrees out of phase with respect to ATP hydrolysis, with the NBDs remaining in close proximity throughout the transport cycle and operating in an asymmetric allosteric manner. The two models are discussed in the light of recent nuclear magnetic resonance and hydrogen-deuterium exchange mass spectrometry analyses of three ABC exporters.
三磷酸腺苷结合盒(ABC)转运体是一个庞大的主要主动转运体超家族,它们是整合膜蛋白,能够将能量与底物跨细胞膜的正向向量转运偶联起来,同时伴随 ATP 的水解。ABC 转运体存在于所有生物体中,主要在原核生物中协调输入,在真核生物中协调输出。与高度保守的核苷酸结合结构域(NBDs)不同,跨膜结构域(TMDs)中的序列保守性较低,其不同的性质可能反映了需要根据质量和极性容纳广泛的底物类型。在过去的十五年中,高分辨率结构分析的爆炸式发展为 ABC 产生了丰富的结构信息。基于这些结构,提出了 ABC 转运体的一般机制,称为开关或交替访问模型,该模型认为 NBDs 是广泛分离的,TMDs 和 NBDs 一起形成一个面向细胞内的倒置“V”形。两个 ATP 和底物与内向构象结合诱导向外向构象的转变。尽管取得了这种明显的进展,但对于任何给定的 ABC,其运输机制的确定性仍然难以捉摸。底物结合和运输与 ATP 结合和水解的偶联方式尚不清楚,并且有大量生化和生物物理数据与广泛分离的 NBDs 作为功能生理状态不一致。已经提出了一种替代的恒接触模型,其中两个 NBS 相对于 ATP 水解以 180 度的相位差操作,在整个运输循环中,NBDs 保持接近,并且以不对称变构方式操作。在最近对三种 ABC 外排泵的核磁共振和氘氢交换质谱分析的基础上,讨论了这两种模型。