Work

A quantitative single-cell assay for retrograde membrane traffic enables rapid detection of defects in cellular organization

Year: 2019

Type: article

Abstract: Retrograde membrane trafficking from plasma membrane to Golgi and endoplasmic reticulum typifies one of the key sorting steps emerging from the early endosome that affects cell surface and intracellular protein dynamics underlying cell function. While some cell surface proteins and lipids are known to sort retrograde, there are few effective methods to quantitatively measure the extent or kinetics of these events. Here we took advantage of the well-known retrograde trafficking of cholera toxin and newly defined split fluorescent protein technology to develop a quantitative, sensitive, and effectively real-time single-cell flow cytometry assay for retrograde membrane transport. The approach can be applied in high throughput to elucidate the underlying biology of membrane traffic and how endosomes adapt to the physiologic needs of different cell types and cell states.

Source: Molecular Biology of the Cell

DOI: 10.1091/mbc.e19-07-0375

Authors: Phi Luong, Qian Li, Pin-Fang Chen, Paul J. Wrighton, Denis Chang, Sean Dwyer, Marie-Theres Bayer, Scott B. Snapper, Steen H. Hansen, Jay R. Thiagarajah, Wolfram Goessling, Wayne I. Lencer

Institutions: Boston Children's Hospital, Harvard University, Shanghai Jiao Tong University, Shanghai Children's Hospital, Brigham and Women's Hospital, Massachusetts General Hospital, Dana-Farber Cancer Institute, Harvard Stem Cell Institute

Language: English

Topic: Cellular transport and secretion

Subfield: Cell Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, National Institute on Alcohol Abuse and Alcoholism, National Cancer Institute, National Institute of Diabetes and Digestive and Kidney Diseases, Eunice Kennedy Shriver National Institute of Child Health and Human Development

Awards: Mechanism of action for the epithelial-specific ER stress sensor IRE1β in regulating intestinal homeostasis and host defense, Training Program in Newborn and Developmental Diseases, Estrogen Regulation of Hepatic Growth, FcRn-dependent sorting of IgG and IgG-opsinized antigens by epithelial cells, The role of mechanical signaling in alcoholic liver disease and hepatocellular carcinoma, p190 RhoGAP signaling in epithelial oncogenesis, Intestinal Disease-enterocyte toxin interaction, Aquaporin 3 dependent responses of colon epithelial cells to microbes and injury, Metabolic Regulation of Liver Growth, Boston Intellectual and Developmental Disabilities Research Center, A Community Zebrafish Resource for Modeling GWAS Biology, Integrated Epithelial and Muscosal Biology, COngenital Diarrhea and Enteropathy (PediCODE) Consortium and BioRepository

OpenAlex ID: W2991127958

FWCI: 0.788

Cites: 55

Cited by: 12

Related to: 10