UREG is a molecular chaperone that works in urea enzyme activation, a nickel-dependent enzyme required by many pathogenic bacteria and fungi to infect their hosts. UREG multitasking also acts as a GTPase enzyme, working with other partners to direct energy from GTP hydrolysis, thereby delivering nickel ions essential for activation of urea enzymes, thus playing a key role in cellular ecosystems and metal homeostasis pathways.
In a landmark study, Dr. Elisabetta Mileo of Bip Laboratory (Aix Marseille University and CNRS) and Professor Barbara Zambelli from the Ministry of Pharmacy and Biotechnology (University of Bologna) and Professor Valérie Belle, Professor Bruno Guigliarelli, Dr. Emilien Etienne Etienne , Dr. Emilien Etienne, Dr. Guillaume Gerbaud, Hugo Leguenno, Ketty Tamburrini and Annalisa Pierro of Aix Marseille University, discovered the dynamic behavior of the GTPase Ureg protein in living cell cells. This represents a significant leap in our understanding of protein function in natural habitats. Their study, published in the Journal of Dear, elucidates how the Ureg protein, essential for activation of bacteria, exhibits significant flexibility in its physiological competition, which may be critical for its enzyme activity.
Previously, the role of UREG as a partner in assisting the activation of nickel-dependent enzymes such as diapers has been well demonstrated by in vitro studies, which demonstrates the flexible behavior of the enzyme in solution. These data promote understanding by combining deployment site-oriented spin labeling with electron paramagnetic resonance (SDSL-EPR) spectroscopy to directly observe the behavior of UREGs in living cells.
Dr. Zambelli emphasized the significance of the study, noting: “The results show that UREG maintains a diverse structural landscape internalexists in the conformational ensemble of two main populations, showing the characteristics of random coil-like or compactness. These insights affirm the physiological significance of the inherent disordered nature of Ureg and propose the effect of protein flexibility on this particular enzyme, which may be related to the regulation of the extensive protein interactions delivered by metal ion. ”
Researchers use innovative technologies to penetrate Ureg protein behavior in cellular environments. “internal EPR experiments were performed by first marking the protein of interest produced by recombinant expression and then passing it in the cell. “Dr Mileo explained. The study involved delivery protocols involving calcium ions and transient heat shock (incubation at 42°C), which promoted optimal protein internalization that retains cell viability.
This study not only highlights the inherent barriers and flexibility of UREG proteins, but also marks a significant methodology in protein research in its natural cellular environment. “Nitrooxide-based spin labels can be performed at room temperature for EPR analysis, compatible with the lifespan of the cells under study.” By leveraging advanced SDSL-EPR spectroscopy, the research team provides the conformational situation of UREGs Invaluable insights that lay the foundation for future efforts aim to uncover the complex mechanisms of protein function in living cells. This study not only deepens our understanding of protein dynamics, but also develops new avenues for new drugs and antimicrobial molecules for the development of targeted therapies and biotechnology applications.
Journal Reference
Annalisa Pierro et al., “Intracellular study of conformational situations of GTPase UREG by SDSL-EPR,” Iscience, 2023.
doi: https://doi.org/10.1016/j.isci.2023.107855.
About the Author
Barbara Zambelli – Associate Professor in 2021, Assistant Professor at Unibo since 2008. Her research activities aim to understand molecular and structural levels to understand, Metal-driven protein interactions In biological systems, this is responsible for regulating metabolic processes that are important to life. In particular, her main research work involves cell metabolism nickel And its counterpart Human healthpossible medical and drug applications.
Elisabetta Mileo He is a CNRS researcher in the laboratory of “Bioénergétiqueeting nierie des Protéines” in Marseille, France. She is a chemist and a spectroscopist with experience and experience in using nitrogen oxide radicals as spin labels in structural biology. E. Mileo research activities focused on the study of protein structural dynamics, especially in chaperones and other flexible proteins, with the aim of obtaining information on how protein dynamics affect protein function. The originality of her work lies in the fact that protein studies of SDSL-EPR are directly inside the cell (internal EPR), under physiological conditions. Her research also aims to develop new tools and new spin tags to follow EPR directly to “action” proteins into live cells.
