Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Odense, Denmark.
Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford, Oxford, UK.
Protein Sci. 2024 Oct;33(10):e5162. doi: 10.1002/pro.5162.
Jumonji-C (JmjC) domain-containing protein 7 (JMJD7) is a human Fe(II) and 2-oxoglutarate dependent oxygenase that catalyzes stereospecific C3-hydroxylation of lysyl-residues in developmentally regulated GTP binding proteins 1 and 2 (DRG1/2). We report studies exploring a diverse set of lysine derivatives incorporated into the DRG1 peptides as potential human JMJD7 substrates and inhibitors. The results indicate that human JMJD7 has a relatively narrow substrate scope beyond lysine compared to some other JmjC hydroxylases and lysine-modifying enzymes. The geometrically constrained (E)-dehydrolysine is an efficient alternative to lysine for JMJD7-catalyzed C3-hydroxylation. γ-Thialysine and γ-azalysine undergo C3-hydroxylation, followed by degradation to formylglycine. JMJD7 also catalyzes the S-oxidation of DRG1-derived peptides possessing methionine and homomethionine residues in place of lysine. Inhibition assays show that DRG1 variants possessing cysteine/selenocysteine instead of the lysine residue efficiently inhibit JMJD7 via cross-linking. The overall results inform on the substrate selectivity and inhibition of human JMJD7, which will help enable the rational design of selective small-molecule and peptidomimetic inhibitors of JMJD7.
Jumonji-C (JmjC) 结构域蛋白 7 (JMJD7) 是一种人类 Fe(II) 和 2-氧戊二酸依赖性氧合酶，可催化发育调节 GTP 结合蛋白 1 和 2 (DRG1/2) 中赖氨酸残基的立体特异性 C3-羟化。我们报告了探索一系列不同的赖氨酸衍生物被整合到 DRG1 肽中作为潜在的人类 JMJD7 底物和抑制剂的研究。结果表明，与其他一些 JmjC 羟化酶和赖氨酸修饰酶相比，人类 JMJD7 的底物范围相对较窄，除了赖氨酸之外。在 JMJD7 催化的 C3-羟化反应中，几何受限的（E）-脱氢赖氨酸是赖氨酸的有效替代物。γ-硫代赖氨酸和γ-氮代赖氨酸发生 C3-羟化，随后降解形成甲酰甘氨酸。JMJD7 还催化具有甲硫氨酸和高半胱氨酸残基而不是赖氨酸残基的 DRG1 衍生肽的 S-氧化。抑制实验表明，具有半胱氨酸/硒代半胱氨酸而不是赖氨酸残基的 DRG1 变体通过交联有效地抑制 JMJD7。总体结果阐明了人类 JMJD7 的底物选择性和抑制作用，这将有助于合理设计 JMJD7 的选择性小分子和肽模拟抑制剂。
