通过时间分辨磷光去极化测量肌质网Ca2+转运三磷酸腺苷酶的节段运动和旋转扩散。

Segmental motion and rotational diffusion of the Ca2+-translocating adenosine triphosphatase of sarcoplasmic reticulum, measured by time-resolved phosphorescence depolarization.

作者信息

Speirs A, Moore C H, Boxer D H, Garland P B

出版信息

Biochem J. 1983 Jul 1;213(1):67-74. doi: 10.1042/bj2130067.

DOI:10.1042/bj2130067

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1152091/

Abstract

We studied the rotational mobility of the Ca2+ + Mg2+-activated ATPase in skeletal-muscle sarcoplasmic-reticulum vesicles, using time-resolved measurements of the depolarization of laser-flash-excited phosphorescence of the extrinsic triplet probe erythrosin. Our results are in general agreement with those of others [Bürkli & Cherry (1981) Biochemistry 20, 138-145] obtained by linear dichroism methods. In addition, we directly observed fast depolarization in the 1-5 microseconds time range that can be attributed to limited motion of part of the protein (segmental motion). Temperature-dependent changes in phosphorescence anisotropy indicated the onset of a conformational change in structure of the Ca2+ + Mg2+-activated ATPase at 11-13 degrees C. We also describe the synthesis of 5-iodoacetamidoerythrosin.

摘要

我们利用对外部三重态探针赤藓红激光闪光激发磷光去极化的时间分辨测量，研究了骨骼肌肌浆网囊泡中Ca2+ + Mg2+激活的ATP酶的旋转流动性。我们的结果与其他人[布尔克利和切里（1981年），《生物化学》20卷，第138 - 145页]通过线性二色性方法得到的结果总体一致。此外，我们直接观察到在1 - 5微秒时间范围内的快速去极化，这可归因于蛋白质部分区域的有限运动（片段运动）。磷光各向异性的温度依赖性变化表明，在11 - 13摄氏度时，Ca2+ + Mg2+激活的ATP酶结构发生了构象变化。我们还描述了5 - 碘乙酰氨基赤藓红的合成。

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3

Fluorescence energy transfer as an indicator of Ca2+-ATPase interactions in sarcoplasmic reticulum.荧光能量转移作为肌浆网中Ca2 + -ATP酶相互作用的指标。

Biophys J. 1987 Feb;51(2):205-20. doi: 10.1016/S0006-3495(87)83326-X.

本文引用的文献

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Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。

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FEBS Lett. 1980 Mar 10;111(2):407-12. doi: 10.1016/0014-5793(80)80838-6.

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Time-dependent absorption anisotropy and rotational diffusion of proteins in membranes.膜中蛋白质的时间依赖性吸收各向异性和旋转扩散

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Anisotropic rotation of bacteriorhodopsin in lipid membranes. Comparison of theory with experiment.细菌视紫红质在脂质膜中的各向异性旋转。理论与实验的比较。

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Biochem J. 1982 Apr 1;203(1):313-21. doi: 10.1042/bj2030313.

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Rotational and lateral movements in biomembranes: the dynamics of biomembrane components.

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Ann N Y Acad Sci. 1981;366:176-96. doi: 10.1111/j.1749-6632.1981.tb20753.x.

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