Slow Dissociation from the PARP1-HPF1 Complex Drives Inhibitor Potency.
Slow Dissociation from the PARP1-HPF1 Complex Drives Inhibitor Potency.
复制标题
DOI：
10.1021/acs.biochem.3c00243
复制
发表时间：
2023-08-15
期刊：
影响因子：
2.9
通讯作者：
Rudolph, Johannes
中科院分区：
文献类型：
--
作者：
Stojanovic, Petra;Luger, Karolin;Rudolph, Johannes
PARP1与受损DNA结合后，被激活对自身和其他蛋白质进行聚adp核糖基化（PARylation），导致染色质松弛和DNA修复因子的募集。HPF1最近被发现是PARP1的一种蛋白辅助因子，通过促进和改变PARP1活性位点，指导组蛋白优先PARP1化。PARP1抑制剂（PARPi）用于治疗BRCA - / -癌症，但其在细胞中的效力基础，特别是在HPF1的背景下，尚不完全清楚。在这里，我们展示了8种不同的PARPi与PARP1的简单一步关联，测量的关联率（kon）为0.8-6 μM-1 s-1。当将PARP1与PARP1 - hpf1复合物进行比较时，我们发现这些比率只有微小的差异。通过表征最近发现的两种PARPi的解离速率（koff）和结合常数（KD），我们发现，例如，saruparib的解离半衰期为22.5 h，而fluzoparib在HPF1存在时对PARP1具有更高的亲和力，就像结构相关的化合物olaparib一样。通过测量KD和kon来计算koff，我们发现细胞中PARPi的效价与PARP1-HPF1复合物的koff相关度最好。我们的数据表明，药物化合物与PARP1-HPF1复合物的解离应该是指导下一代PARPi开发的选择参数。
PARP1, upon binding to damaged DNA, is activated to perform poly ADP-ribosylation (PARylation) on itself and other proteins, which leads to relaxation of chromatin and recruitment of DNA repair factors. HPF1 was recently discovered as a protein cofactor of PARP1 that directs preferential PARylation of histones over other targets by contributing to and altering the PARP1 active site. Inhibitors of PARP1 (PARPi) are used in the treatment of BRCA–/– cancers, but the basis for their potency in cells, especially in the context of HPF1, is not fully understood. Here, we demonstrate the simple one-step association for eight different PARPi to PARP1 with measured rates of association (kon) of 0.8–6 μM–1 s–1. We find only minor differences in these on rates when comparing PARP1 with the PARP1–HPF1 complex. By characterizing the rates of dissociation (koff) and the binding constants (KD) for two more recently discovered PARPi, we find, for example, that saruparib has a half-life for dissociation of 22.5 h and fluzoparib has higher affinity for PARP1 in the presence of HPF1, just like the structurally related compound olaparib. By using the measured KD and kon to calculate koff, we find that the potency of PARPi in cells correlates best with the koff from the PARP1–HPF1 complex. Our data suggest that dissociation of a drug compound from the PARP1–HPF1 complex should be the parameter of choice for guiding the development of next-generation PARPi.
登录
查看更多内容
HPF1 重塑 PARP1 的活性位点，以实现组蛋白的丝氨酸 ADP 核糖基化。
影响因子：
16.6
作者：
Sun FH;Zhao P;Zhang N;Kong LL;Wong CCL;Yun CH
通讯作者：
Yun CH
影响因子：
16
作者：
Gibbs-Seymour I;Fontana P;Rack JGM;Ahel I
通讯作者：
Ahel I
影响因子：
7.3
作者：
Menear, Keith A.;Adcock, Claire;Martin, Niall M. B.
通讯作者：
Martin, Niall M. B.
影响因子：
11.5
作者：
Donawho, Cherrie K.;Luo, Yan;Frost, David J.
通讯作者：
Frost, David J.
影响因子：
64.8
作者：
Scully, R;Livingston, DM
通讯作者：
Livingston, DM