Specific cardiolipin binding interferes with labeling of sulfhydryl residues in the adenosine diphosphate/adenosine triphosphate carrier protein from beef heart mitochondria
Specific cardiolipin binding interferes with labeling of sulfhydryl residues in the adenosine diphosphate/adenosine triphosphate carrier protein from beef heart mitochondria
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DOI：
10.1021/bi9610055
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发表时间：
1996-12-10
期刊：
影响因子：
2.9
通讯作者：
Nuscher, B
中科院分区：
文献类型：
--
作者：
Beyer, K;Nuscher, B
本文研究了牛心线粒体中分离的ADP/ATP载体蛋白与心磷脂的相互作用，揭示了伴随该蛋白构象转变的一个半胱氨酸基残基Cys56。23、107-113]。用巯基试剂N-(1-吡喃基)马来酰亚胺(PYM)的静态荧光监测揭幕过程。在心磷脂(CL)存在下，ADP启动构象转变后观察到的平阳霉素结合率显著降低。除CL外，其他磷脂的效果要差得多。结果表明，CL对构象转变和结合反应均有影响，但影响程度不同。磷脂酶Az消化蛋白结合的磷脂后，依赖于ADP的PYM结合率增加。磷脂酶处理还增加了不依赖ADP的PYM结合，从而表明ADP对载体转变的控制逐渐丧失。如果磷脂酶孵育在大约1小时后终止，则通过添加CL可以完全恢复ADP的控制。这些结果将与早期关于心磷脂紧密结合的报道[Beyer，K.，&Klingenberg，M.(1985)BioChemical 24,3821-3826]和当前的ADP/ATP载体蛋白的结构模型进行讨论。
The interaction of cardiolipin with the isolated ADP/ATP carrier protein from beef heart mitochondria has been studied by means of the unmasking of a single cysteinyl residue, Cys56, which accompanies the conformational transition of the protein [Leblanc, P., & Clauser, H. (1972) FEES Lett. 23, 107-113]. The unmasking was monitored by using the static fluorescence of the sulfhydryl reagent N-(1-pyrenyl)maleimide (PYM). The rate of PYM binding that was observed after initiation of the conformational transition by ADP was drastically reduced in the presence of cardiolipin (CL). Phospholipids other than CL were much less effective. It can be shown that the conformational transition and the binding reaction are both affected by CL, although to varying extents. An enhancement of the rate of the ADP-dependent PYM binding was observed upon digestion of the protein bound phospholipid by phospholipase Az. The phospholipase treatment also led to an increased ADP-independent PYM binding, thus indicating that the ADP control of the carrier transition was gradually lost. The ADP control could be fully restored through the addition of CL, provided that the phospholipase incubation had been terminated after approximately 1 h. These results will be discussed in relation to an earlier report of tight cardiolipin binding [Beyer, K., & Klingenberg, M. (1985) Biochemistry 24, 3821-3826] and to current structural models of the ADP/ATP carrier protein.