Expedient Synthesis of Ubiquitin‐like Protein ISG15 Tools through Chemo‐Enzymatic Ligation Catalyzed by a Viral Protease Lbpro

Ubiquitin (Ub)‐like protein ISG15 (interferon‐stimulated gene 15) regulates innate immunity and links with the evasion of host response by viruses such as SARS‐CoV‐2. Dissecting ISGylation pathways recently received increasing attention which can inform related disease interventions, but such studies necessitate the preparation and development of various ISG15 protein tools. Here, we find that the leader protease (Lbpro) encoded by foot‐and‐mouth disease virus can promote ligation reactions between recombinant ISG15 and synthetic glycyl compounds, generating protein tools such as ISG15‐propargylamide and ISG15‐rhodamine110, which are needed for cellular proteomic studies of deISGylases, and the screening and evaluation of inhibitors against SARS‐CoV‐2 papain‐like protease (PLpro). Furthermore, this strategy can be also used to load ISG15 onto the lysine of a synthetic peptide through an isopeptide bond, and prepare Ub and NEDD8 (ubiquitin‐like protein Nedd8) protein tools.

Expedient Synthesis of Ubiquitin-like Protein ISG15 Tools through Chemo-Enzymatic Ligation Catalyzed by a Viral Protease Lbpro.

Ubiquitin (Ub)-like protein ISG15 (interferon-stimulated gene 15) regulates innate immunity and links with the evasion of host response by viruses such as SARS-CoV-2. Dissecting ISGylation pathways recently received increasing attention which can inform related disease interventions, but such studies necessitate the preparation and development of various ISG15 protein tools. Here, we find that the leader protease (Lbpro ) encoded by foot-and-mouth disease virus can promote ligation reactions between recombinant ISG15 and synthetic glycyl compounds, generating protein tools such as ISG15-propargylamide and ISG15-rhodamine110, which are needed for cellular proteomic studies of deISGylases, and the screening and evaluation of inhibitors against SARS-CoV-2 papain-like protease (PLpro). Furthermore, this strategy can be also used to load ISG15 onto the lysine of a synthetic peptide through an isopeptide bond, and prepare Ub and NEDD8 (ubiquitin-like protein Nedd8) protein tools.

Evolving from Laboratory Toys towards Life-Savers: Small-Scale Magnetic Robotic Systems with Medical Imaging Modalities.

Small-scale magnetic robots are remotely actuated and controlled by an externally applied magnetic field. These robots have a characteristic size ranging from several millimetres down to a few nanometres. They are often untethered in order to access constrained and hard-to-reach space buried deep in human body. Thus, they promise to bring revolutionary improvement to minimally invasive diagnostics and therapeutics. However, existing research is still mostly limited to scenarios in over-simplified laboratory environment with unrealistic working conditions. Further advancement of this field demands researchers to consider complex unstructured biological workspace. In order to deliver its promised potentials, next-generation small-scale magnetic robotic systems need to address the constraints and meet the demands of real-world clinical tasks. In particular, integrating medical imaging modalities into the robotic systems is a critical step in their evolution from laboratory toys towards potential life-savers. This review discusses the recent efforts made in this direction to push small-scale magnetic robots towards genuine biomedical applications. This review examines the accomplishment achieved so far and sheds light on the open challenges. It is hoped that this review can offer a perspective on how next-generation robotic systems can not only effectively integrate medical imaging methods, but also take full advantage of the imaging equipments to enable additional functionalities.