Brooks J C, Brooks M
Life Sci. 1985 Nov 18;37(20):1869-75. doi: 10.1016/0024-3205(85)90003-7.
Permeabilized cells treated with the adenosine triphosphate analog, [35S]adenosine-5'-0-(3-thiotriphosphate) ([gamma-35S]ATP), showed thiophosphorylation of a small number of cellular proteins. A 54 kilodalton (kDa) protein was heavily thiophosphorylated in unstimulated control cells and a 43 kilodalton protein was more heavily thiophosphorylated in calcium stimulated cells. Intact cells incorporated 35S into a series of higher molecular weight proteins. Stimulation of prelabelled, permeabilized cells resulted in a loss of 35S from the cells over a 20 min period. Treatment of permeabilized cells with ATP gamma S inhibited secretion and 35S incorporation into the cells. Pretreatment with ATP gamma S resulted in subsequent inhibition of both secretion and the ability of the cells to incorporate 35S from [gamma-35S]ATP. These results indicate that the sites normally available for phosphorylation were inactivated by thiophosphorylation and were unavailable to participate in the secretory process. The inhibition of secretion associated with thiophosphorylation of these proteins suggests that they may play a role in the control of secretion by chromaffin cells.
用三磷酸腺苷类似物[35S]腺苷-5'-O-(3-硫代三磷酸)([γ-35S]ATP)处理的透化细胞显示出少量细胞蛋白的硫代磷酸化。在未刺激的对照细胞中，一种54千道尔顿(kDa)的蛋白被大量硫代磷酸化，而在钙刺激的细胞中，一种43千道尔顿的蛋白被更大量地硫代磷酸化。完整细胞将35S掺入一系列更高分子量的蛋白质中。对预先标记的透化细胞进行刺激导致在20分钟内细胞中35S的损失。用ATPγS处理透化细胞会抑制分泌以及35S掺入细胞。用ATPγS预处理会导致随后分泌以及细胞从[γ-35S]ATP掺入35S的能力均受到抑制。这些结果表明，正常可用于磷酸化的位点因硫代磷酸化而失活，无法参与分泌过程。与这些蛋白的硫代磷酸化相关的分泌抑制表明它们可能在嗜铬细胞分泌的控制中起作用。
