期刊
CHEMICAL SCIENCE
卷 12, 期 36, 页码 12181-12191出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d1sc04065k
关键词
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资金
- ARAID
- Spanish Ministry of Science, Innovation and Universities [BFU2016-75633-P, PID2019-105451GB-I00, PID2019-109395GB-I00]
- Gobierno de Aragon [E34_R17, LMP58_18]
- FEDER
- Biotechnology and Biological Sciences Research Council (BBSRC) [BB/P010660/1, BB/M011216/1]
- Universidad de Sevilla (Acciones Especiales del VI Plan Propio de Investigacion y Transferencia)
- National Institute of Allergy and Infectious Diseases (NIAID) [AI127973, AI153202]
- National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health [P20GM130448]
NleB/SseK effectors modify host cell proteins through Tyr284 residue to regulate substrate selectivity, ultimately affecting bacterial virulence in a host-specific manner.
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 Tyr284(NleB/NleB1), a second-shell residue contiguous to the catalytic machinery. Tyr284(NleB/NleB1) is important in coupling protein substrate binding to catalysis. This is exemplified by S286Y(SseK1) and N302Y(SseK2) 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.
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