Neurotransporter Group, Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, 0317, Oslo, Norway.
Departamento de Biología Molecular, Universidad Autónoma de Madrid, Madrid, Spain.
Neurochem Res. 2022 Jan;47(1):85-110. doi: 10.1007/s11064-021-03331-z. Epub 2021 Apr 27.
In contrast to water soluble enzymes which can be purified and studied while in solution, studies of solute carrier (transporter) proteins require both that the protein of interest is situated in a phospholipid membrane and that this membrane forms a closed compartment. An additional challenge to the study of transporter proteins has been that the transport depends on the transmembrane electrochemical gradients. Baruch I. Kanner understood this early on and first developed techniques for studying plasma membrane vesicles. This advanced the field in that the experimenter could control the electrochemical gradients. Kanner, however, did not stop there, but started to solubilize the membranes so that the transporter proteins were taken out of their natural environment. In order to study them, Kanner then had to find a way to reconstitute them (reinsert them into phospholipid membranes). The scope of the present review is both to describe the reconstitution method in full detail as that has never been done, and also to reveal the scientific impact that this method has had. Kanner's later work is not reviewed here although that also deserves a review because it too has had a huge impact.
与可在溶液中进行纯化和研究的水溶性酶不同,溶质载体(转运蛋白)的研究既要求感兴趣的蛋白质位于磷脂膜中,又要求该膜形成封闭隔室。转运蛋白研究的另一个挑战是,运输取决于跨膜电化学梯度。Baruch I. Kanner 很早就理解了这一点,并首先开发了用于研究质膜小泡的技术。这使得实验者可以控制电化学梯度,从而推动了该领域的发展。然而,Kanner 并没有就此止步,而是开始溶解膜,以便将转运蛋白从其自然环境中取出。为了研究它们,Kanner 必须找到一种重新构建它们的方法(重新插入磷脂膜)。本综述的范围不仅详细描述了这种重建方法,因为这从未做过,而且还揭示了这种方法的科学影响。此处未回顾 Kanner 的后期工作,尽管这也值得回顾,因为它也产生了巨大的影响。