Improved Electrophoretic Separation to Assist the Monitoring of Bcl-xL Post-Translational Modifications
by
Claude Bobo 1,2, Claire Céré 1,2, Mélody Dufossée 1,2, Alain Dautant 1,2
, Violaine Moreau 3,4, Stéphen Manon 1,2, Florian Beaumatin 1,2 and Muriel Priault 1,2,*
Claude Bobo 1,2, Claire Céré 1,2, Mélody Dufossée 1,2, Alain Dautant 1,2, Violaine Moreau 3,4, Stéphen Manon 1,2, Florian Beaumatin 1,2 and Muriel Priault 1,2,*
1
CNRS, Institut de Biochimie et de Génétique Cellulaires, UMR 5095, 1 Rue Camille Saint-Saëns, 33077 Bordeaux, France
2
Université de Bordeaux, Institut de Biochimie et de Génétique Cellulaires, UMR5095, 1 Rue Camille Saint-Saëns, 33077 Bordeaux, France
3
INSERM U1053, Bordeaux Research in Translational Oncology, 146, Rue Léo Saignat, 33076 Bordeaux, France
4
Université de Bordeaux, 146, Rue Léo Saignat, 33076 Bordeaux, France
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(22), 5571; https://doi.org/10.3390/ijms20225571
Received: 26 July 2019 / Revised: 25 October 2019 / Accepted: 5 November 2019 / Published: 8 November 2019
(This article belongs to the Special Issue Advances in Bcl-xL Research)
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.
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MDPI and ACS Style
Bobo, C.; Céré, C.; Dufossée, M.; Dautant, A.; Moreau, V.; Manon, S.; Beaumatin, F.; Priault, M. Improved Electrophoretic Separation to Assist the Monitoring of Bcl-xL Post-Translational Modifications. Int. J. Mol. Sci. 2019, 20, 5571.
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