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Monofunctionalization with dextrans endows cell‐penetrative properties to proteins
Author(s) -
Kilgore Henry Ralph,
Ressler Valerie T,
Raines Ronald T
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.634.7
Subject(s) - cytochrome c , dextran , apoptosome , cytochrome , chemistry , biochemistry , apoptosis , cytosol , microbiology and biotechnology , caspase , biophysics , biology , programmed cell death , enzyme
Dextrans have proven to be a widely useful class of polysaccharides, with applications ranging from antithrombotic agents to enhancers of vaccine stability. We have examined an unprecedented property of dextrans: the capacity to convey cargo into the cytosol. Mono‐functionalized dextran–flourogenic probe and dextran–protein conjugates were prepared to assess the ability of dextran to mediate cellular delivery. We show that dextran conjugates are taken up in an energy‐dependent process culminating in cytosolic access. Several cell types were examined, indicating the generality of this uptake pathway. These observations motivated the preparation of dextran–cytochrome C conjugates. Release of cytochrome C from the intermembrane space activates the intrinsic apoptosis pathway. Cytotoxicity measurements of dextran–cytochrome C conjugates thus provide a straightforward metric for cytosolic access of dextran–protein conjugates. We verify that cytochrome C mediates cell death by measuring activation of downstream caspases 3 and 7, indicating that dextran–cytochrome C conjugates promote formation of the apoptosome. These experiments document a new and exciting frontier for polysaccharide bioconjugates and support the existence of an uptake pathway for 1,6‐linked polysaccharides. Support or Funding Information [This work was supported by Grant R01 GM044783 (NIH).] This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .