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The Tick-Over Theory Revisited: Formation and Regulation of the Soluble Alternative Complement C3 Convertase (C3(H2O)Bb)

机译：翻阅理论：可溶性替代补体C3转化酶（C3（H2O）Bb）的形成和调控

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摘要

The molecular interactions between the components of the C3 convertase of the alternative pathway (AP) of complement and its regulators, in both surface-bound and fluid-phase form, are still incompletely understood. The fact that the AP convertase is labile makes studies difficult to perform. According to the so called tick-over theory, hydrolyzed C3, called C3(H2O), forms the initial convertase in fluid phase together with factor B. In the present study, we have applied western blot analysis and ELISA together with fluorescence resonance energy transfer (FRET) to study the formation of the fluid-phase AP convertases C3(H2O)Bb and C3bBb and their regulation by factor H and factor I at specific time points and, with FRET, in real time. In our hands, factor B showed a higher affinity for C3(H2O) than for C3b, although in both cases it was readily activated to Bb. However, the convertase activity of C3bBb was approximately twice that of C3(H2O)Bb, as monitored by the generation of C3a. But in contrast, the C3(H2O)Bb convertase was more resistant to inactivation by factor H and factor I than was the C3bBb convertase. Under conditions that totally inactivated C3bBb, C3(H2O)Bb still retained approximately 25% of its initial activity.
机译：补体的替代途径（AP）的C3转化酶的成分与其调节剂之间的分子相互作用，无论是表面结合形式还是液相形式，都尚未完全了解。 AP转化酶不稳定的事实使得研究难以进行。根据所谓的滴答理论，水解后的C3（称为C3（H2O））与因子B一起在液相中形成初始转化酶。在本研究中，我们将Western blot分析和ELISA与荧光共振能量转移一起应用（FRET）研究液相AP转化酶C3（H2O）Bb和C3bBb的形成及其在特定时间点受H因子和I因子的调节，以及FRET实时研究。在我们手中，因子B对C3（H2O）的亲和力高于对C3b的亲和力，尽管在两种情况下它都易于活化为Bb。但是，C3bBb的转化酶活性大约是C3（H2O）Bb的两倍，这是通过C3a的产生来监测的。但是相比之下，C3（H2O）Bb转化酶比H3bBb转化酶对H因子和I因子的灭活更有抵抗力。在完全灭活C3bBb的条件下，C3（H2O）Bb仍保留约25％的初始活性。

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