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Aonbangkhen, Chanat

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Aonbangkhen

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Aonbangkhen, Chanat

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    Target Protein Deglycosylation in Living Cells by a Nanobody-Fused Split O-GlcNAcase
    (Springer Science and Business Media LLC, 2021-03-08) Ge, Yun; Ramirez, Daniel H.; Yang, Bo; D’Souza, Alexandria K.; Aonbangkhen, Chanat; Wong, Stephanie; Woo, Christina M.
    O-GlcNAc is an essential and dynamic post-translational modification presented on thousands of nucleocytoplasmic proteins. Interrogating the role of O-GlcNAc on a single target protein is crucial yet challenging to perform in cells. Herein, we developed a nanobody-fused split O-GlcNAcase (OGA) as an O-GlcNAc eraser for selective deglycosylation of a target protein in cells. After systematic cellular optimization, we identified a split OGA with reduced inherent deglycosidase activity that selectively removed O-GlcNAc from the desired target protein when directed by a nanobody. We demonstrate the generality of the nanobody-fused split OGA using four nanobodies against five target proteins, and use the system to study functions for O-GlcNAc on the transcription factors c-Jun and c-Fos. The nanobody-directed O-GlcNAc eraser provides a new strategy for functional evaluation and engineering of O-GlcNAc via the selective removal of O-GlcNAc from individual proteins directly in cells.