Synthetic retinals: convenient probes of rhodopsin and visual transduction process.

Synthetic retinals: convenient probes of rhodopsin and visual transduction process.
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合成视网膜:视紫红质和视觉转导过程的便捷探针。

DOI:
10.1016/s0076-6879(00)15846-x
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发表时间:
2000
影响因子:
--
通讯作者:
Crouch,RK
Crouch,RK
中科院分区:
生物学4区
文献类型:
--
作者:
Lou,J;Tan,Q;Karnaukhova,E;Berova,N;Nakanishi,K;Crouch,RK

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All the visual pigment proteins are integral membrane proteins, crossing the membrane seven times with the majority of the protein being composed of hydrophobic amino acids within the membrane. This has made these proteins difficult to study by many standard biophysical techniques. However, each of these opsins has a chromophore, generally ll-cis-retinal or one of a few closely related structures, that is attached to the protein via a protonated Schiff base linkage. The site of attachment is at a lysine in the center of the seventh helix. In vitro, exposure of these proteins to light in the presence of hydroxylamine results in the complete detachment of the retinal, and washing the membranes with bovine serum albumin (1%) removes any residual retinal. This generates the opsin apoprotein, which can reform rhodopsin on the addition of ll-cis-retinal in the dark. This provides a most convenient tool for the examination of rhodopsin structure and function as the protein has also been found to accommodate various retinal isomers and analogs, forming pigments or complexes which can then be tested for functionality and physical properties. A large number of retinals have been synthesized and tested with opsins from various species (see a recent review1). Most of these experiments have been carried out in vitro although a few studies have shown that retinal analogs can be successfully incorporated into some species including vitamin A-deficient rats. 2 Very little work to date has been conducted on the cone opsins due to problems in obtaining reasonable quantities of the proteins. However, with the development of both improved expression systems and purification techniques, these studies are now being initiated on the cone opsins as well. Interestingly, the physiologic recordings conducted to date on isolated rod and cone photoreceptors utilizing retinal analogs indicate that there are some significant differences in the interactions of these respective opsins with their chromophores. 3 Recently studies have been initiated
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