Improved Electrophoretic Separation to Assist the Monitoring of Bcl-xL Post-Translational Modifications

• Published in: International journal of molecular sciences Vol. 20; no. 22; p. 5571
• Main Authors: Bobo, Claude, Céré, Claire, Dufossée, Mélody, Dautant, Alain, Moreau, Violaine, Manon, Stéphen, Beaumatin, Florian, Priault, Muriel
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 08.11.2019; MDPI
• Subjects: Apoptosis; bcl-X Protein - metabolism; Electrophoresis - methods; HCT116 Cells; Humans; Life Sciences; Molecular weight; Mutant Proteins - isolation & purification; Mutation - genetics; Phosphorylation; Protein Processing, Post-Translational; Proteins; deamidation; Bcl-xL; post-translational modification; electrophoresis; phosphorylation; Bcl-x L
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20225571

Bcl-xL 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.