1. Improved Electrophoretic Separation to Assist the Monitoring of Bcl-xL Post-Translational Modifications.
- Author
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Bobo, Claude, Céré, Claire, Dufossée, Mélody, Dautant, Alain, Moreau, Violaine, Manon, Stéphen, Beaumatin, Florian, and Priault, Muriel
- Subjects
EXERGONIC reactions ,BCL-2 proteins ,DEAMINATION ,PHARMACEUTICAL gels - Abstract
Bcl-x
L is an oncogene of which the survival functions are finely tuned by post-translational modifications (PTM). Within the Bcl-2 family of proteins, Bcl-xL shows unique eligibility to deamidation, a time-related spontaneous reaction. Deamidation is still a largely overlooked PTM due to a lack of easy techniques to monitor Asn→Asp/IsoAsp conversions or Glu→Gln conversions. Being able to detect PTMs is essential to achieve a comprehensive description of all the regulatory mechanisms and functions a protein can carry out. Here, we report a gel composition improving the electrophoretic separation of deamidated forms of Bcl-xL generated either by mutagenesis or by alkaline treatment. Importantly, this new gel formulation proved efficient to provide the long-sought evidence that even doubly-deamidated Bcl-xL remains eligible for regulation by phosphorylation. [ABSTRACT FROM AUTHOR]- Published
- 2019
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