Tracking Chaperone-Mediated Autophagy Flux with a pH-Resistant Fluorescent Reporter DOI Open Access
Robert Z. Qi, Xianzhong Chen, Zihan Li

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 26(1), С. 17 - 17

Опубликована: Дек. 24, 2024

Chaperone-mediated autophagy (CMA) is a selective autophagic pathway responsible for degrading cytoplasmic proteins within lysosomes. Monitoring CMA flux essential understanding its functions and molecular mechanisms but remains technically complex challenging. In this study, we developed pH-resistant probe, KFERQ-Gamillus, by screening various green fluorescent proteins. This probe activated under conditions known to induce CMA, such as serum starvation, relies on LAMP2A the KFERQ motif lysosomal localization degradation, demonstrating specificity pathway. It enables detection of activity in living cells through both microscopy image-based flow cytometry. Additionally, created dual-reporter system, KFERQ-Gamillus-Halo, integrating KFERQ-Gamillus with Halo-tag system. not only distinguishes between protein synthesis degradation also facilitates intracellular via immunoblotting rapid assessment using Together, KFERQ-Gamillus-Halo provides quantitative time-resolved monitoring cells. tool holds promising potential high-throughput biomedical research related CMA.

Язык: Английский

The link between ferroptosis and autophagy in myocardial ischemia/reperfusion injury: new directions for therapy DOI
Xiaoting Yang, Hui Wu,

Di Liu

и другие.

Journal of Cardiovascular Translational Research, Год журнала: 2025, Номер unknown

Опубликована: Янв. 30, 2025

Язык: Английский

Процитировано

1
Sex-specific and cell-type-specific changes in chaperone-mediated autophagy across tissues during aging DOI Creative Commons
Rabia R. Khawaja, Adrián Martín‐Segura, Olaya Santiago‐Fernández

и другие.

Nature Aging, Год журнала: 2025, Номер unknown

Опубликована: Фев. 5, 2025

Aging leads to progressive decline in organ and tissue integrity function, partly due loss of proteostasis autophagy malfunctioning. A decrease with age chaperone-mediated (CMA), a selective type lysosomal degradation, has been reported various organs cells from rodents humans. Disruption CMA recapitulates features aging, whereas activating mice protects against age-related diseases such as Alzheimer's, retinal degeneration and/or atherosclerosis. However, sex-specific cell-type-specific differences aging remain unexplored. Here, using reporter single-cell transcriptomic data, we report that most cell types show age, males exhibiting greater aging. Reduced is often associated fewer lysosomes competent for CMA. Transcriptional downregulation genes may further contribute decline, especially males. These findings suggest influence vulnerability degeneration. Using imaging fluorescent (CMA) RNA sequencing the authors present resource on basal activity across organs, sexes young old mice, offering comprehensive overview changes this proteostatic mechanism context

Язык: Английский

Процитировано

1
T-Cell Senescence: Unlocking the Tumor Immune “Dark Box” - A Multidimensional Analysis from Mechanism to Tumor Immunotherapeutic Intervention DOI
Jia Cheng, Jian Zheng, Man Chen

и другие.

Seminars in Cancer Biology, Год журнала: 2025, Номер unknown

Опубликована: Май 1, 2025

Язык: Английский

Процитировано

0
Tracking Chaperone-Mediated Autophagy Flux with a pH-Resistant Fluorescent Reporter DOI Open Access
Robert Z. Qi, Xianzhong Chen, Zihan Li

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 26(1), С. 17 - 17

Опубликована: Дек. 24, 2024

Chaperone-mediated autophagy (CMA) is a selective autophagic pathway responsible for degrading cytoplasmic proteins within lysosomes. Monitoring CMA flux essential understanding its functions and molecular mechanisms but remains technically complex challenging. In this study, we developed pH-resistant probe, KFERQ-Gamillus, by screening various green fluorescent proteins. This probe activated under conditions known to induce CMA, such as serum starvation, relies on LAMP2A the KFERQ motif lysosomal localization degradation, demonstrating specificity pathway. It enables detection of activity in living cells through both microscopy image-based flow cytometry. Additionally, created dual-reporter system, KFERQ-Gamillus-Halo, integrating KFERQ-Gamillus with Halo-tag system. not only distinguishes between protein synthesis degradation also facilitates intracellular via immunoblotting rapid assessment using Together, KFERQ-Gamillus-Halo provides quantitative time-resolved monitoring cells. tool holds promising potential high-throughput biomedical research related CMA.

Язык: Английский

Процитировано

0