A micro-batchwise technique method for rapid reconstitution of functionally active mitochondrial ADP/ATP carrier from Jerusalem artichoke (Helianthus tuberosus L.) tubers

Journal of biochemical and biophysical methods Pub Date : 2008-04-24 DOI:10.1016/j.jprot.2007.11.003

A. Spagnoletta , A. De Palma , G. Prezioso , V. Scalera

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引用次数: 9

Abstract

A method for rapid reconstitution of ADP/ATP carrier from Jerusalem artichoke (Helianthus tuberosus L.) tubers mitochondria in proteoliposomes is described. The method is based on the well known property of the Amberlite resin to absorb the detergent allowing proteoliposome formation. This has been achieved by a micro-batchwise technique, using a rotating plate stirrer. An evaluation of the optimal conditions, in comparison with the more usual column method is presented. The purified ADP/ATP carrier, incorporated in proteoliposomes by this method, shows a high transport activity and a higher specific activity with respect to proteoliposomes obtained by the column procedure. Furthermore the proteoliposomal preparations are more homogeneous in size, with a diameter ranging from 300 to 350 nm. The method is suitable for the reconstitution of other membrane transport proteins.

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微批量快速重建菊芋块茎中功能活性线粒体ADP/ATP载体的方法

本文报道了一种从菊芋块茎蛋白脂质体线粒体中快速重建ADP/ATP载体的方法。该方法是基于众所周知的特性，安伯利特树脂吸收洗涤剂，允许形成蛋白脂质体。这是通过使用旋转板搅拌器的微批量技术实现的。对最优条件进行了评价，并与常用的柱法进行了比较。通过这种方法将纯化的ADP/ATP载体结合到蛋白脂质体中，与柱法获得的蛋白脂质体相比，显示出较高的运输活性和更高的比活性。此外，蛋白脂质体制剂在尺寸上更均匀，直径范围为300至350纳米。该方法也适用于其他膜转运蛋白的重构。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Journal of biochemical and biophysical methods

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