Evidence for the presence of disulfide bridges in opioid receptors essential for ligand binding. Possible role in receptor activation
Evidence for the presence of disulfide bridges in opioid receptors essential for ligand binding. Possible role in receptor activation
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阿片受体中存在配体结合所必需的二硫键的证据。
DOI：
--
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发表时间：
1989
影响因子：
2.7
通讯作者：
E. J. Simon
中科院分区：
文献类型：
--
作者：
T. Gioannini;Liu Yf;Park Yh;J. Hiller;E. J. Simon
纯化的μ阿片结合蛋白在SDS-聚丙烯酰胺凝胶电泳中的迁移率对还原剂的存在敏感。在DTT浓度增加的情况下，表观分子量以逐步方式从53 kDa增加至65 kDa。这种迁移率的降低归因于二硫键的连续断裂，导致分子越来越不对称。用还原剂（如DTT）处理来自不同脑区的细胞膜，对阿片类药物结合产生浓度依赖性抑制。对DTT抑制的敏感性在受体类型之间变化，μ > δ κ。对于μ受体，激动剂结合对DTT比拮抗剂结合敏感得多。DTT的抑制作用很容易逆转，并且不受Na+和/或Mg 2+离子的影响。通过加入低浓度的还原剂（如谷胱甘肽）可部分防止可逆性，还原剂不会抑制结合，但会阻断二硫键的重新形成。饱和度数据的Scatchard分析表明，DTT导致结合亲和力的显着降低，对受体数量的影响很小。有人建议，二硫键是必不可少的配体结合和切割的一个或多个这些债券可能发挥作用，阿片受体激动剂的激活。
The mobility of purified μ opioid binding protein in SDS‐polyacrylamide gek electrophoresis is sensitive to the presence of reducing agents. In the presence of increasing concentrations of DTT the apparent molecular weight increases in a stepwise fashion from 53 kDa to 65 kDa. This reduction in mobility is attributed to the successive breakage of disulfide bridges, resulting in an increasingly asymmetric molecule. Treatment of cell membranes from various brain areas with reducing agents, such as DTT, produced a concentration‐dependent inhibition of opioid binding. Sensitivity to DTT inhibition varied between receptor types, μ > δ ≫ κ. For μ receptors, agonist binding was considerably more sensitive to DTT than antagonist binding. Inhibition by DTT is readily reversible and is unaffected by Na+ and/or Mg2+ ions. Reversibility may be partially prevented by the inclusion of a low concentration of a reducing reagent such as glutathione which does not inhibit binding but blocks reformation of disulfide bonds. Scatchard analysis of saturation data shows that DTT causes a pronounced decrease in binding affinity with little effect on receptor number. It is suggested that disulfide bonds are essential for ligand binding and that cleavage of one or more of these bonds may play a role in opioid receptor activation by agonists.
在存在或不存在激动剂的情况下，硫醇对 β-肾上腺素能受体的功能激活。
DOI：
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发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pedersen,SE;Ross,EM
通讯作者：
Ross,EM
大鼠心脏α1-肾上腺素能受体的光亲和标记。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Terman,BI;Insel,PA
通讯作者：
Insel,PA