Department of Chemistry, University of Pennsylvania, Philadelphia, PA, 19104, USA.
Abramson Family Cancer Research Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
Nat Commun. 2020 Feb 10;11(1):818. doi: 10.1038/s41467-020-14584-7.
The human N-terminal acetyltransferase E (NatE) contains NAA10 and NAA50 catalytic, and NAA15 auxiliary subunits and associates with HYPK, a protein with intrinsic NAA10 inhibitory activity. NatE co-translationally acetylates the N-terminus of half the proteome to mediate diverse biological processes, including protein half-life, localization, and interaction. The molecular basis for how NatE and HYPK cooperate is unknown. Here, we report the cryo-EM structures of human NatE and NatE/HYPK complexes and associated biochemistry. We reveal that NAA50 and HYPK exhibit negative cooperative binding to NAA15 in vitro and in human cells by inducing NAA15 shifts in opposing directions. NAA50 and HYPK each contribute to NAA10 activity inhibition through structural alteration of the NAA10 substrate-binding site. NAA50 activity is increased through NAA15 tethering, but is inhibited by HYPK through structural alteration of the NatE substrate-binding site. These studies reveal the molecular basis for coordinated N-terminal acetylation by NatE and HYPK.
人类 N-端乙酰转移酶 E（NatE）包含 NAA10 和 NAA50 催化亚基以及 NAA15 辅助亚基，并与具有内在 NAA10 抑制活性的 HYPK 蛋白相关联。NatE 共翻译乙酰化了大约一半的蛋白质组，以介导多种生物学过程，包括蛋白质半衰期、定位和相互作用。NatE 和 HYPK 如何合作的分子基础尚不清楚。在这里，我们报告了人类 NatE 和 NatE/HYPK 复合物的冷冻电镜结构以及相关的生物化学研究。我们揭示了 NAA50 和 HYPK 通过诱导 NAA15 向相反方向移动，在体外和人体细胞中表现出对 NAA15 的负协同结合。NAA50 和 HYPK 都通过改变 NAA10 底物结合位点的结构来抑制 NAA10 活性。通过与 NAA15 的连接增加 NAA50 的活性，但通过改变 NatE 底物结合位点的结构来抑制 HYPK 的活性。这些研究揭示了 NatE 和 HYPK 协调 N 端乙酰化的分子基础。
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