Label-free protein quantification of sodium butyrate treated CHO cells by ESI-UHR-TOF-MS

- Limited knowledge of molecular productivity enhancing effects of butyrate on CHO.
- Quick impact of butyrate on histone modification visible by AU-PAGE Western blotting.
- Straight forward label-free MS-quantification improved view on shifted cellular state.
- Altered protein level in cell growth, carbohydrate metabolism, protein expression.
- HDAC inhibition by butyrate also visible in changes of nucleosome modifying factors.

Effects of butyrate on CHO producer cells are contradictory, promoting productivity and at the same time repressing proliferation. Though in previous omics studies the background of butyrate impact on producer cells has been investigated, the knowledge about the mechanism is still very limited. As previous proteomic results on this field are mainly based on 2DE-gels, we conducted a label-free MS quantification, based on fast high resolution ESI-MS and a straight forward software solution, to gain insight in shifted cellular processes of CHO cells 25 h after butyrate treatment. 118 proteins or subunits with significantly altered abundances were identified suggesting changes in carbohydrate, protein metabolic and cell cycle processes. Effects of butyrate on the nucleosome assembly as a known direct epigenetic influence on HDAC activity turned out to be unexpectedly fast and persistent, as confirmed by Western blots of histone-H4 acetylation. Contradictory to increased cell specific productivity, most elements of protein metabolism exhibited decreased levels after butyrate treatment. In comparison to published results some overlap of our label free MS data could be observed but also apparently diverging findings, showing the need for complementary omics techniques for a holistic view on cellular processes such as response to butyrate.