通过光漂白后双光子荧光恢复测量全反式视黄醇的流动性。
Measurement of the mobility of all-trans-retinol with two-photon fluorescence recovery after photobleaching.
作者信息
Koutalos Yiannis
机构信息
Departments of Ophthalmology and Neurosciences, Medical University of South Carolina, Charleston, SC 29425, USA.
出版信息
Methods Mol Biol. 2010;652:115-27. doi: 10.1007/978-1-60327-325-1_6.
The mobility of all-trans-retinol makes a crucial contribution to the rate of the reactions in which it participates. This is even more so because of its low aqueous solubility, which makes the presence of carrier proteins and the spatial arrangement of cellular membranes especially relevant. In rod photoreceptor outer segments, all-trans-retinol is generated after light exposure from the reduction of all-trans-retinal that is released from bleached rhodopsin. The mobility of all-trans-retinol in rod outer segments was measured with fluorescence recovery after photobleaching (FRAP), using two-photon excitation of its fluorescence. The values of the lateral and axial diffusion coefficients indicate that most of the all-trans-retinol in rod outer segments move unrestricted and without being aided by carriers.
全反式视黄醇的流动性对其参与的反应速率起着至关重要的作用。由于其低水溶性，使得载体蛋白的存在以及细胞膜的空间排列显得尤为重要，情况更是如此。在视杆光感受器外段，全反式视黄醇是在光照后由漂白视紫红质释放的全反式视黄醛还原产生的。利用双光子激发其荧光，通过光漂白后荧光恢复（FRAP）技术测量了全反式视黄醇在视杆外段的流动性。横向和轴向扩散系数的值表明，视杆外段中的大多数全反式视黄醇不受限制地移动，且无需载体协助。
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