Etemadi A H
Biochim Biophys Acta. 1980 Dec 31;604(3):347-422. doi: 10.1016/0005-2736(80)90578-7.
This and the companion article are aimed at surveying the methods used for the study of membrane asymmetry. The techniques employed for the assessment of the asymmetric distribution and orientation of membrane proteins are reviewed in this article, whereas those pertaining to the unequal distribution of lipids are detailed in the companion paper. The use of immunological techniques and lectins, functions of proteins and their perturbations, chemical reagents, enzymatic isotopic labeling and enzymatic cleavage of membrane proteins and physical techniques are discussed and illustrated using recent examples of their application. Whenever appropriate, problems involving crypticity and non-availability or non-reactivity of functional sites, relevant chemical functions or protein fragments to appropriate ligands, reagents or modifying enzymes are envisaged and possible modification of the exposure of proteins during preparation of ghosts and other drawbacks are discussed, the use of different techniques and control experiments in conjunction is recommended for a more realistic assessment of the distribution and orientation of proteins.
本文与配套文章旨在综述用于研究膜不对称性的方法。本文回顾了用于评估膜蛋白不对称分布和取向的技术，而配套论文则详细阐述了与脂质不均等分布相关的技术。本文讨论并通过近期应用实例说明了免疫技术和凝集素的使用、蛋白质的功能及其扰动、化学试剂、酶促同位素标记以及膜蛋白的酶促裂解和物理技术。在适当的时候，设想了涉及隐蔽性以及功能位点、相关化学功能或蛋白质片段对合适配体、试剂或修饰酶不可用或无反应性的问题，并讨论了在制备血影和其他缺点过程中蛋白质暴露的可能改变，建议结合使用不同技术和对照实验，以便更实际地评估蛋白质的分布和取向。
