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Multiple post-translational modifications affect heterologous protein synthesis
Alexander A Tokmakov1, Atsushi Kurotani, Tetsuo Takagi
1RIKEN Systems and Structural Biology Center, University of Tokyo, Bunkyo, Tokyo 113-0033, Japan. tokmak@phoenix.kobe-u.ac.jp
The Journal of Biological Chemistry
|June 8, 2012
Summary
Post-translational modifications (PTMs) impact protein production. Some PTMs hinder soluble expression, while others like phosphorylation enhance it, aiding recombinant protein synthesis and stability.
Area of Science:
- Biochemistry and Molecular Biology
- Protein Engineering
- Recombinant Protein Expression
Background:
- Post-translational modifications (PTMs) are crucial for protein function and folding.
- Prokaryotic systems have limited capacity for PTMs, hindering heterologous protein production.
- A systematic analysis of individual PTM effects on heterologous protein synthesis is lacking.
Purpose of the Study:
- To investigate the correlation between predicted post-translational modification sites and the expression yield of heterologous proteins.
- To identify specific PTMs that influence the solubility and stability of recombinant proteins.
- To provide insights for optimizing heterologous protein synthesis.
Main Methods:
- Expression of 1488 human proteins and domains in a bacterial cell-free system.
- Bioinformatic prediction of multiple post-translational modification sites.
- Statistical analysis of correlations between predicted PTMs and protein expression yields.
Main Results:
- Myristoylation, glycosylation, palmitoylation, and disulfide bond formation negatively correlated with soluble protein expression.
- Aspartyl hydroxylation, C-terminal amidation, and Tyr sulfation showed no significant correlation with expression yield.
- Phosphorylation, ubiquitination, SUMOylation, and prenylation were associated with increased production of soluble, folded proteins.
Conclusions:
- Predicted PTMs can serve as indicators for heterologous protein expression success.
- Understanding PTMs' roles can guide strategies for optimizing recombinant protein synthesis.
- This study offers significant evidence for the impact of PTMs on recombinant protein stability and solubility.
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