Department of Pharmacy, University of Patras, 26504, Patras, Greece.
Biomol NMR Assign. 2022 Oct;16(2):399-406. doi: 10.1007/s12104-022-10110-6. Epub 2022 Sep 15.
hPARP14 is a human ADP-ribosyl-transferase (ART) that belongs to the macroPARPs family, together with hPARP9 and hPARP15. It contains a tandem of three macro domains (MD) while each of them has different properties. The first one, namely MD1, has not been reported to exhibit a high binding affinity for ADP-ribose (ADPr) in contrast to the following two (MD2 and MD3). All three MDs exhibit an α/β/α sandwich-like fold as reported by the deposited crystallographic structures. MD2 and MD3 recognize mono-ADP-ribosylated (MARylated) but not poly-ADP-ribosylated (PARylated) substrates and thus they allow hPARP14 to bind its targets, which can be potentially MARylated by its catalytic domain (CD). hPARP14 participates in DNA damage repair process and immune response against viruses like SARS-CoV-2, which also harbors an MD fold. Furthermore, hPARP14 like the other two macroPARPs (hPARP9 and hPARP15), is implicated in numerous types of cancer, such as B-aggressive lymphoma and sarcoma, rendering its MDs as potential important drug targets. Herein, we report the complete NMR backbone and side chain assignment (H, C, N) of hPARP14 MD2 in the free and ADPr bound states and the NMR chemical shift-based prediction of its secondary structure elements. This is the first reported NMR study of a hPARP macro domain, paving the way to screen by NMR chemical compounds which may alter the ability of hPARP14 to interact with its substrates affecting its function.
hPARP14 是一种人类 ADP-ribosyl-transferase（ART），属于宏 PARP 家族，与 hPARP9 和 hPARP15 一起。它包含三个串联的宏结构域（MD），而每个 MD 都具有不同的特性。第一个，即 MD1，与后两个（MD2 和 MD3）不同，它没有报道显示对 ADP-ribose（ADPr）具有高结合亲和力。所有三个 MD 都表现出由沉积的晶体结构报告的 α/β/α 三明治样折叠。MD2 和 MD3 识别单 ADP-ribosylated（MARylated）但不识别多 ADP-ribosylated（PARylated）底物，因此它们允许 hPARP14 与其靶标结合，其催化结构域（CD）可以潜在地将其 MARylated。hPARP14 参与 DNA 损伤修复过程和对 SARS-CoV-2 等病毒的免疫反应，SARS-CoV-2 也具有 MD 折叠。此外，hPARP14 与其他两种宏 PARP（hPARP9 和 hPARP15）一样，与多种癌症有关，如 B 侵袭性淋巴瘤和肉瘤，使其 MD 成为潜在的重要药物靶标。本文报道了 hPARP14 MD2 在游离和 ADPr 结合状态下的完整 NMR 骨架和侧链分配（H、C、N）以及其二级结构元件的基于 NMR 化学位移的预测。这是 hPARP 宏结构域的首次报道的 NMR 研究，为通过 NMR 化学化合物筛选铺平了道路，这些化合物可能改变 hPARP14 与底物相互作用的能力，从而影响其功能。
