Girdlestone J, Bisson R, Capaldi R A
Biochemistry. 1981 Jan 6;20(1):152-6. doi: 10.1021/bi00504a025.
The interaction of purified succinate dehydrogenase and succinate--ubiquinone reductase (complex II) with lipids was explored by using two (arylazido)phospholipids, one with the reactive nitrene in the head-group region of the bilayer [1-palmitoyl-2-(2-azido-4-nitrobenzoyl)-sn-glycero-3-[3H]phosphocholine (PLII)] and one with the nitrene on the methyl terminus of one of the fatty acid chains [1-myristoyl-2-[12-[(2-azido-4-nitrophenyl)amino]lauroyl]-sn-glycero-3-[14C]phosphocholine (PLI)]. Protein was reacted with vesicles of egg lecithin containing radioactive (arylazido)-phospholipids and the covalent cross-linking of lipid and protein induced by irradiation under UV light. Purified succinate dehydrogenase was found to bind to lipid vesicles through both subunits as both were labeled by PLII. The smaller subunit was inserted into the interior of the bilayer and labeled by PLI. Complex II was found to interact with lipid vesicles, with the smaller subunit of succinate dehydrogenase, CII-3, and CII-4 all inserted into the interior of the bilayer. The large subunit of succinate dehydrogenase was found to be held above the bilayer in complex II and not labeled by either probe. Results are used to derive a picture of the arrangement of subunits in complex II and to evaluate the utility of (arylazido)-phospholipids in membrane studies.
通过使用两种（芳基叠氮）磷脂来探究纯化的琥珀酸脱氢酶和琥珀酸 - 泛醌还原酶（复合体II）与脂质的相互作用，一种磷脂的反应性氮烯位于双层的头部区域[1 - 棕榈酰 - 2 - （2 - 叠氮基 - 4 - 硝基苯甲酰基） - sn - 甘油 - 3 - [3H]磷酸胆碱（PLII）]，另一种磷脂的氮烯位于其中一条脂肪酸链的甲基末端[1 - 肉豆蔻酰 - 2 - [12 - [（2 - 叠氮基 - 4 - 苯基）氨基]月桂酰基] - sn - 甘油 - 3 - [14C]磷酸胆碱（PLI）]。使蛋白质与含有放射性（芳基叠氮）磷脂的卵磷脂囊泡反应，并通过紫外光照射诱导脂质与蛋白质的共价交联。发现纯化的琥珀酸脱氢酶通过两个亚基与脂质囊泡结合，因为两个亚基都被PLII标记。较小的亚基插入双层内部并被PLI标记。发现复合体II与脂质囊泡相互作用，琥珀酸脱氢酶的较小亚基CII - 3和CII - 4都插入双层内部。发现琥珀酸脱氢酶的大亚基在复合体II中位于双层上方，并且未被任何一种探针标记。研究结果用于推导复合体II中亚基排列的图像，并评估（芳基叠氮）磷脂在膜研究中的实用性。
