NleB/SseK-catalyzed arginine-glycosylation and enteropathogen virulence are finely tuned by a single variable position contiguous to the catalytic machinery.
NleB/SseK-catalyzed arginine-glycosylation and enteropathogen virulence are finely tuned by a single variable position contiguous to the catalytic machinery.
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DOI：
10.1039/d1sc04065k
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发表时间：
2021-09-22
期刊：
影响因子：
8.4
通讯作者：
Hurtado-Guerrero R
中科院分区：
文献类型：
--
作者：
García-García A;Hicks T;El Qaidi S;Zhu C;Hardwidge PR;Angulo J;Hurtado-Guerrero R
NleB/SseK效应子是由肠道细菌病原体表达的甘氨酸-GlcNAc-转移酶，其修饰宿主细胞蛋白以破坏信号传导途径。而保守的啮齿柠檬酸杆菌NleB和E.大肠杆菌NleB 1蛋白对宿主蛋白具有广泛的选择性，沙门氏菌SseK 1、SseK 2和SseK 3对宿主蛋白具有狭窄的蛋白底物选择性。在这里，通过结合计算和生物物理实验，我们证明了广泛的蛋白质底物选择性NleB依赖于Tyr 284 NleB/NleB 1，第二壳残基相邻的催化机器。Tyr 284 NleB/NleB 1在偶联蛋白质底物结合催化中是重要的。S286 YSseK 1和N302 YSseK 2突变体就是例证，它们分别对FADD和DR 3死亡结构域具有活性，并且其动力学性质与肠出血性大肠杆菌的动力学性质相匹配。coli NleB1。将这些突变体整合到S. enterica增加S.肠杆菌在巨噬细胞中的存活，表明更好的酶动力学参数导致增强的毒力。我们的研究结果提供了深入了解这些酶如何精细地调节甘氨酸糖基化，进而调节细菌毒力。此外，我们的数据显示如何混杂糖基转移酶优先糖基化特定的蛋白质底物。NleB和SseK糖基转移酶以不同的底物特异性糖基化哺乳动物蛋白质的精氨酸残基。我们发现，这些差异依赖于一个特定的第二壳残基邻近的催化机器。
NleB/SseK effectors are arginine-GlcNAc-transferases expressed by enteric bacterial pathogens that modify host cell proteins to disrupt signaling pathways. While the conserved Citrobacter rodentium NleB and E. coli NleB1 proteins display a broad selectivity towards host proteins, Salmonella enterica SseK1, SseK2, and SseK3 have a narrowed protein substrate selectivity. Here, by combining computational and biophysical experiments, we demonstrate that the broad protein substrate selectivity of NleB relies on Tyr284NleB/NleB1, a second-shell residue contiguous to the catalytic machinery. Tyr284NleB/NleB1 is important in coupling protein substrate binding to catalysis. This is exemplified by S286YSseK1 and N302YSseK2 mutants, which become active towards FADD and DR3 death domains, respectively, and whose kinetic properties match those of enterohemorrhagic E. coli NleB1. The integration of these mutants into S. enterica increases S. enterica survival in macrophages, suggesting that better enzymatic kinetic parameters lead to enhanced virulence. Our findings provide insights into how these enzymes finely tune arginine-glycosylation and, in turn, bacterial virulence. In addition, our data show how promiscuous glycosyltransferases preferentially glycosylate specific protein substrates. The NleB and SseK glycosyltransferases glycosylate arginine residues of mammalian proteins with different substrate specificities. We uncover that these differences rely on a particular second-shell residue contiguous to the catalytic machinery.
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影响因子：
16.6
作者：
Lira-Navarrete, Erandi;de las Rivas, Matilde;Companon, Ismael;Carmen Pallares, Maria;Kong, Yun;Iglesias-Fernandez, Javier;Bernardes, Goncalo J. L.;Peregrina, Jesus M.;Rovira, Carme;Bernado, Pau;Bruscolini, Pierpaolo;Clausen, Henrik;Lostao, Anabel;Corzana, Francisco;Hurtado-Guerrero, Ramon
通讯作者：
Hurtado-Guerrero, Ramon
影响因子：
16.6
作者：
de Las Rivas M;Lira-Navarrete E;Daniel EJP;Compañón I;Coelho H;Diniz A;Jiménez-Barbero J;Peregrina JM;Clausen H;Corzana F;Marcelo F;Jiménez-Osés G;Gerken TA;Hurtado-Guerrero R
通讯作者：
Hurtado-Guerrero R
影响因子：
4.6
作者：
El Qaidi, Samir;Scott, Nichollas E.;Hardwidge, Philip R.
通讯作者：
Hardwidge, Philip R.
影响因子：
5.7
作者：
Gan, Jiyao;Scott, Nichollas E.;Giogha, Cristina
通讯作者：
Giogha, Cristina
影响因子：
30.3
作者：
Gao X;Wang X;Pham TH;Feuerbacher LA;Lubos ML;Huang M;Olsen R;Mushegian A;Slawson C;Hardwidge PR
通讯作者：
Hardwidge PR