Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling ...Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling is activated by integrins and interferons,which can induce ISG15 gene expression.This gene encodes a ubiquitin-like protein that functions as a protein modifier via the ISGylation system.Furthermore,non-conjugated ISG15 acts as a cytokine-like protein.In this viewpoint,the interplay between free ISG15,protein ISGylation,and cytoskeletal dynamics in the tumor microenvironment is discussed.展开更多
Interferon-stimulated gene 15(ISG15)and its associated post-translational modification,ISGylation,have evolved from being a mere antiviral effector to a central regulator of diverse cellular processes[1].Beyond its ca...Interferon-stimulated gene 15(ISG15)and its associated post-translational modification,ISGylation,have evolved from being a mere antiviral effector to a central regulator of diverse cellular processes[1].Beyond its canonical role in immune responses,ISGylation—the covalent attachment of ISG15 to substrates—modulates lysosomal function,autophagy,and redox balance.Recent studies reveal its unexpected involvement in copper metabolism,a pathway critical for both cellular homeostasis and pathogen defense.Copper,while essential as a cofactor for enzymes like cytochrome C oxidase and superoxide dismutase(SOD1),and involved in numerous biological processes,is toxic when excess,triggering cuproptosis—a copper-dependent mitochondrial cell death[2,3].Understanding the interplay between ISG15/ISGylation and copper-related pathways can provide insights into fundamental processes of life.展开更多
ISG15 is a ubiquitin-like(Ubl) protein attached to substrate proteins by ISG15 conjugating enzymes whose dysregulation is implicated in a multitude of disease processes, but the probing of these enzymes remains to be ...ISG15 is a ubiquitin-like(Ubl) protein attached to substrate proteins by ISG15 conjugating enzymes whose dysregulation is implicated in a multitude of disease processes, but the probing of these enzymes remains to be accomplished. Here, we describe the development of a new activity-based probe ISG15-Dha(dehydroalanine) through protein semi-synthesis. In vitro crosslinking and cell lysate proteomic profiling experiments showed that this probe can sequentially capture ISG15 conjugating enzymes including E1 enzyme UBA7, E2 enzyme UBE2L6, E3 enzyme HERC5, the previously known ISG15 deconjugating enzyme(USP18), as well as some other enzymes(USP5 and USP14) which we additionally confirmed to impart deISGylation activity. Collectively, ISG15-Dha provides a new tool that can simultaneously capture ISG15 conjugating and deconjugating enzymes for biochemical or pharmacological studies.展开更多
基金This research was supported by the School of Science and Technology(CCyT-2021-5)from the Autonomous University of Mexico City(UACM).
文摘Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling is activated by integrins and interferons,which can induce ISG15 gene expression.This gene encodes a ubiquitin-like protein that functions as a protein modifier via the ISGylation system.Furthermore,non-conjugated ISG15 acts as a cytokine-like protein.In this viewpoint,the interplay between free ISG15,protein ISGylation,and cytoskeletal dynamics in the tumor microenvironment is discussed.
基金supported by the Natural Science Foundation of China(grant number 82211530059 to J.X.).
文摘Interferon-stimulated gene 15(ISG15)and its associated post-translational modification,ISGylation,have evolved from being a mere antiviral effector to a central regulator of diverse cellular processes[1].Beyond its canonical role in immune responses,ISGylation—the covalent attachment of ISG15 to substrates—modulates lysosomal function,autophagy,and redox balance.Recent studies reveal its unexpected involvement in copper metabolism,a pathway critical for both cellular homeostasis and pathogen defense.Copper,while essential as a cofactor for enzymes like cytochrome C oxidase and superoxide dismutase(SOD1),and involved in numerous biological processes,is toxic when excess,triggering cuproptosis—a copper-dependent mitochondrial cell death[2,3].Understanding the interplay between ISG15/ISGylation and copper-related pathways can provide insights into fundamental processes of life.
基金supported by the National Key Research&Development Program of China(2021YFA1200104,2022YFC3401500)the National Natural Science Foundation of China(21621003,22137005,21971133,22027807,22034004,92253302,22227810)the Tsinghua University Spring Breeze Fund(2020Z99CFY043,2021Z99CFZ002)。
文摘ISG15 is a ubiquitin-like(Ubl) protein attached to substrate proteins by ISG15 conjugating enzymes whose dysregulation is implicated in a multitude of disease processes, but the probing of these enzymes remains to be accomplished. Here, we describe the development of a new activity-based probe ISG15-Dha(dehydroalanine) through protein semi-synthesis. In vitro crosslinking and cell lysate proteomic profiling experiments showed that this probe can sequentially capture ISG15 conjugating enzymes including E1 enzyme UBA7, E2 enzyme UBE2L6, E3 enzyme HERC5, the previously known ISG15 deconjugating enzyme(USP18), as well as some other enzymes(USP5 and USP14) which we additionally confirmed to impart deISGylation activity. Collectively, ISG15-Dha provides a new tool that can simultaneously capture ISG15 conjugating and deconjugating enzymes for biochemical or pharmacological studies.
