Evaluation of disulfide reduction during receptor-mediated endocytosis by using FRET imaging

Jun Yang, Hongtao Chen, Iontcho R. Vlahov, Ji Xin Cheng, Philip S. Low*

*Corresponding author for this work

Research output: Contribution to journalArticle

238 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)13872-13877
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number37
DOIs
StatePublished - Sep 12 2006

Keywords

  • Disulfide bond reduction
  • Drug targeting
  • Endosome
  • Folate receptor

ASJC Scopus subject areas

  • General

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