Static and time-resolved fluorescence studies of fluorescent phosphatidylcholine bound to the phosphatidylcholine transfer protein of bovine liver.
Static and time-resolved fluorescence studies of fluorescent phosphatidylcholine bound to the phosphatidylcholine transfer protein of bovine liver.
复制标题
荧光磷脂酰胆碱与牛肝脏磷脂酰胆碱转移蛋白结合的静态和时间分辨荧光研究。
作者：
T. Berkhout;A. Visser;K. Wirtz
磷脂酰胆碱类似物在sn-1、sn-2或sn-1和sn-2位置（分别为1-PnA-PC、2-PnA-PC和diPnA - pc）含有顺式- parinaryl链，通过荧光技术研究了牛肝脏磷脂酰胆碱转移蛋白（PC-TP）的脂质结合位点。这些荧光脂与蛋白质的结合是通过测量parinaroyl荧光增强和色氨酸荧光猝灭来记录的。与PC-TP结合的1-PnA-、2-PnA-和diPnA - pc的荧光强度与发色团的含量成正比。色氨酸残基与结合的发色团之间的能量转移效率在1-PnA-和2-PnA-PC中约为40%，在diPnA - pc中约为60%。色氨酸荧光猝灭的部分原因是荧光寿命的减少。通过时间分辨荧光各向异性测量分析了PnA-PC类似物在转移蛋白上的1和2脂肪酸基链的取向。荧光各向异性根据单一指数函数衰减，1-PnA-PC的旋转相关时间为26 ns， 2-PnA-PC为11 ns， diPnA -PC为15 ns。这些相关时间表明两个脂肪酰基链固定在蛋白质上的不同位置。根据相关时间的差异，我们提出磷脂酰胆碱转移蛋白的形状为椭球状（轴比等于2.5），1脂肪酸链平行于长对称轴，与2脂肪酸链成60-90度角。
Phosphatidylcholine analogues containing a cis- parinaroyl chain at the sn-1, sn-2, or both sn-1 and sn-2 positions (1-PnA-PC, 2-PnA-PC, and diPnA -PC, respectively) have been used to investigate the lipid binding site of the phosphatidylcholine transfer protein (PC-TP) from bovine liver by fluorometric techniques. Binding of these fluorescent lipids to the protein was registered by measuring the enhancement of parinaroyl fluorescence and the quenching of the tryptophanyl fluorescence. The fluorescence intensity of 1-PnA-, 2-PnA- and diPnA -PC bound to PC-TP was proportional to the chromophore content. The energy-transfer efficiency between the tryptophan residues and the bound chromophores was approximately 40% for 1-PnA- and 2-PnA-PC and 60% for diPnA -PC. Quenching of the tryptophanyl fluorescence was, in part, accounted for by a decrease of the fluorescence lifetimes. The orientation of the 1 and 2 fatty acyl chains of the PnA-PC analogues on the transfer protein was analyzed by time-resolved fluorescence anisotropy measurements. The fluorescence anisotropy decayed according to a single exponential function yielding a rotational correlation time of 26 ns for 1-PnA-PC, 11 ns for 2-PnA-PC, and 15 ns for diPnA -PC. These correlation times indicate that both fatty acyl chains are immobilized at different positions on the protein. From the difference in correlation time we propose that the shape of the phosphatidylcholine transfer protein is ellipsoidal (axial ratio congruent to 2.5) with the 1 fatty acyl chain oriented parallel to the long symmetry axis and having an angle of 60-90 degrees with the 2 fatty acyl chain.
二氢吲嗪蛋白与细菌荧光素酶之间的关联：二氢吲嗪发射各向异性衰减的直接证明。
影响因子：
2.9
作者：
Visser,AJ;Lee,J
通讯作者：
Lee,J
大鼠肝脏质膜中的脂质结构域。
DOI：
10.1111/j.1432-1033.1983.tb07391.x
发表时间：
1983
期刊：
European journal of biochemistry
影响因子：
--
作者：
Schroeder,F
通讯作者：
Schroeder,F
鉴定磷脂酰胆碱转移蛋白与含有光活化卡宾前体的磷脂酰胆碱类似物的脂质结合位点。
DOI：
10.1111/j.1432-1033.1983.tb07382.x
发表时间：
1983
期刊：
European journal of biochemistry
影响因子：
--
作者：
Westerman,J;Wirtz,KW;Berkhout,T;vanDeenen,LL;Radhakrishnan,R;Khorana,HG
通讯作者：
Khorana,HG