Hanson P I, Meyer T, Stryer L, Schulman H
Department of Neurobiology, Stanford University School of Medicine, California 94305-5401.
Neuron. 1994 May;12(5):943-56. doi: 10.1016/0896-6273(94)90306-9.
Autophosphorylation of multifunctional Ca2+/calmodulin-dependent protein kinase makes it Ca2+ independent by trapping bound calmodulin and by enabling the kinase to remain partially active even after calmodulin dissociates. We show that autophosphorylation is an intersubunit reaction between neighbors in the multimeric kinase which requires two molecules of calmodulin. Ca2+/calmodulin acts not only to activate the "kinase" subunit but also to present effectively the "substrate" subunit for autophosphorylation. Conversion of the kinase to the potentiated or trapped state is a cooperative process that is inefficient at low occupancy of calmodulin. Simulations show that repetitive Ca2+ pulses at limiting calmodulin lead to the recruitment of calmodulin to the holoenzyme, which further stimulates autophosphorylation and trapping. This cooperative, positive feedback loop will potentiate the response of the kinase to sequential Ca2+ transients and establish a threshold frequency at which the enzyme becomes highly active.
多功能钙/钙调蛋白依赖性蛋白激酶的自磷酸化通过捕获结合的钙调蛋白以及使激酶即使在钙调蛋白解离后仍能保持部分活性,从而使其不依赖于钙。我们发现自磷酸化是多聚体激酶中相邻亚基之间的一种反应,这需要两分子的钙调蛋白。钙/钙调蛋白不仅作用于激活“激酶”亚基,还能有效地呈现“底物”亚基以供自磷酸化。激酶向增强或捕获状态的转变是一个协同过程,在钙调蛋白占有率较低时效率不高。模拟结果表明,在有限的钙调蛋白条件下,重复的钙脉冲会导致钙调蛋白被募集到全酶中,这进一步刺激自磷酸化和捕获。这种协同的正反馈回路将增强激酶对连续钙瞬变的反应,并建立一个阈值频率,在该频率下酶变得高度活跃。