Chemical modification of transducin with dansyl chloride hinders its binding to light-activated rhodopsin.

Chemical modification of transducin with dansyl chloride hinders its binding to light-activated rhodopsin.
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用丹磺酰氯对转导蛋白进行化学修饰会阻碍其与光激活视紫红质的结合。

DOI:
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发表时间:
2004
期刊:
Journal of biochemistry and molecular biology
影响因子:
--
通讯作者:
J. Bubis
J. Bubis
中科院分区:
--
文献类型:
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作者:
A. Kosoy;C. Möller;D. Perdomo;J. Bubis

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转导蛋白(Transducin,T)是视杆细胞外节的异源三聚体鸟嘌呤核苷酸结合蛋白,在受体蛋白视紫红质和效应蛋白cGMP磷酸二酯酶之间起中介作用。用丹磺酰氯(DnsCl)标记T抑制其光依赖性鸟嘌呤核苷酸结合活性。相反,DnsCl对视紫红质的功能没有影响。每摩尔T掺入约2-3摩尔DnsCl。由于氟铝酸盐能够激活DnsCl修饰的T,这种赖氨酸特异性标记化合物不影响T的鸟嘌呤核苷酸结合口袋。然而,标记的T与DnsCl阻碍其结合光激发的视紫红质,如沉降实验所示。此外,视紫红质完全保护T.这些结果证明了T上存在功能性赖氨酸,其位于与光感受器蛋白的相互作用位点附近。
Transducin (T), the heterotrimeric guanine nucleotide binding protein in rod outer segments, serves as an intermediary between the receptor protein, rhodopsin, and the effector protein, cGMP phosphodiesterase. Labeling of T with dansyl chloride (DnsCl) inhibited its light-dependent guanine nucleotide binding activity. Conversely, DnsCl had no effect on the functionality of rhodopsin. Approximately 2-3 mol of DnsCl were incorporated per mole of T. Since fluoroaluminate was capable of activating DnsCl-modified T, this lysine-specific labeling compound did not affect the guanine nucleotide-binding pocket of T. However, the labeling of T with DnsCl hindered its binding to photoexcited rhodopsin, as shown by sedimentation experiments. Additionally, rhodopsin completely protected against the DnsCl inactivation of T. These results demonstrated the existence of functional lysines on T that are located in the proximity of the interaction site with the photoreceptor protein.
转导蛋白功能界面的突变分析。
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DOI: --
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