Labeling of bovine heart cytochrome c oxidase with analogues of phospholipids. Synthesis and reactivity of a new cardiolipin benzaldehyde probe.
Labeling of bovine heart cytochrome c oxidase with analogues of phospholipids. Synthesis and reactivity of a new cardiolipin benzaldehyde probe.
复制标题
用磷脂类似物标记牛心细胞色素 C 氧化酶。
DOI：
10.1021/bi00398a024
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发表时间：
1987
期刊：
影响因子：
2.9
通讯作者：
Griffith,OH
中科院分区：
文献类型：
--
作者：
Kuppe,A;Mrsny,RJ;Shimizu,M;Firsan,SJ;Keana,JF;Griffith,OH
本文报道了心磷脂和磷脂酰胆碱两种新的放射性探针的合成。这些探针是naturallipids的衍生物，并且在头基区域中含有胺特异性苯甲醛。该官能团允许选择反应时间（例如，平衡和去污剂去除后），因为仅在添加还原剂后形成不可逆共价键。这些探针，以及磷脂酸的苯甲醛类似物，和水溶性苯甲醛试剂共价连接到牛心细胞色素c氧化酶。重组成囊泡后，脂质-苯甲醛探针选择性地掺入酶的较小多肽中，而其余亚基（I-IV）几乎没有掺入标记。在这里使用的条件下标记的胺基团的可及性是独立的苯甲醛探针之间的结构和电荷差异。这表明所有三种脂质探针与细胞色素c氧化酶复合物的多肽在膜磷脂的一般接触位点反应。一种水溶性苯甲醛试剂，主要标记亚基IV、Va和Vb以及VII-VIII的多肽。这些结果的比较有利于一个更精细的观点，相对于脂质和水相的细胞色素c氧化酶的多肽的处置。
Revised Manuscript Received May 8, 1987 abstract: The syntheses of two new radioactive probes derived from cardiolipin and phosphatidylcholine are reported. These probes are derivatives of naturallipids and contain an amine-specific benzaldehyde in the head-group region. This functional group allows a choice of timing of the reaction (eg, after equilibration and detergent removal) because an irreversible covalent bond is formed only upon the addition of reducing agent. These probes, as well as a benzaldehyde analogue of phosphatidic acid, and a water-soluble benzaldehyde reagent were covalently attached to bovineheart cytochrome c oxidase. After reconstitution into vesicles, the lipid-benzaldehyde probes selectively incorporated into the smaller polypeptides of the enzyme, while the remaining subunits (I-IV) exhibited little incorporation of label. The accessibility of amine groups labeled under the conditions used here was independent of the structural and charge differences between the benzaldehyde probes. This suggests that all three lipid probes react with polypeptides of the cytochrome c oxidase complex at general contact sites for membrane phospholipids. A water-soluble benzaldehyde reagent predominantly labeled subunits IV, Va, and Vb and polypeptides of VII-VIII. A comparison of these results facilitates a more refined view of the disposition of polypeptides of cytochrome c oxidase in respect to the lipid and aqueous phases.