Evaluation of disulfide reduction during receptor-mediated endocytosis by using FRET imaging
- 12 September 2006
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 103 (37), 13872-13877
- https://doi.org/10.1073/pnas.0601455103
Abstract
Despite functional evidence for disulfide bond-reducing activity in endosomal compartments, the mechanistic details pertaining to such process (e.g., kinetics and sites of disulfide reduction) remain largely controversial. To address these questions directly, we have synthesized a previously uncharacterized fluorescent folate conjugate, folate-(BODIPY FL)-SS-rhodamine (folate-FRET), that changes fluorescence from red to green upon disulfide bond reduction. Using this construct, we have observed that disulfide reduction: (i) occurs with a half-time of 6 h after folate-FRET endocytosis, (ii) begins in endosomes and does not depend significantly on redox machinery located on the cell surface or within the lysosome or the Golgi apparatus, (iii) occurs independently of endocytic vesicle trafficking along microtubules, and (iv) yields products that are subsequently sorted into distinct endosomes and trafficked in different directions. Finally, colocalization of folate and transferrin receptors suggest that conclusions derived from this study may apply to other endocytic pathways.This publication has 43 references indexed in Scilit:
- Identification of a ferrireductase required for efficient transferrin-dependent iron uptake in erythroid cellsNature Genetics, 2005
- Synthesis and Biological Evaluation of EC72: A New Folate-Targeted ChemotherapeuticBioconjugate Chemistry, 2005
- Folate receptor endocytosis and traffickingAdvanced Drug Delivery Reviews, 2004
- A review of folate receptor alpha cycling and 5-methyltetrahydrofolate accumulation with an emphasis on cell models in vitroAdvanced Drug Delivery Reviews, 2004
- Folate receptor-mediated targeting of therapeutic and imaging agents to activated macrophages in rheumatoid arthritisAdvanced Drug Delivery Reviews, 2004
- Gemtuzumab ozogamicin in the treatment of acute myeloid leukemiaCancer, 2003
- Mismatch Binding Protein-Based Mutation Detection SystemsMethods, 1995
- Dissecting the Mechanism of Protein Disulfide Isomerase: Catalysis of Disulfide Bond Formation in a Model PeptideBiochemistry, 1994
- Oxidized Redox State of Glutathione in the Endoplasmic ReticulumScience, 1992
- Effect of pH on the conformation of diphtheria toxin and its implications for membrane penetrationBiochemistry, 1985