| 研究実績の概要 |
This was the third and the last year of the study. The bioinformatics analysis of eukaryotic cell-free expression has been performed. The focus was set upon the connection between intrinsic disorder of amino acid sequences and expression success of cell-free protein synthesis. Recent biochemical and structural data indicate that more than half of eukaryotic proteins possess long intrinsically disordered regions. Cell-free synthesis of these sequences was sought to be rationalized. The analysis revealed that elevated disorder content is associated with the increased ratio of soluble expression, whereas overall propensity for detectable protein expression decreases with disorder content. These tendencies are rooted in the distinct features of intrinsically disordered regions, such as low hydrophobicity, elevated surface accessibility and high abundance of sequence motifs for proteolytic degradation.
Altogether, in the course of this project (FY 2013-2015), a bioinformatics algorithm aimed at identification of multiple physicochemical and structural properties associated with soluble expression of eukaryotic proteins in cell-free extracts has been developed and successfully applied. A number of significant correlations between calculated and predicted properties of amino acid sequences and their propensity for soluble cell-free expression have been revealed. They are of practical use for predicting expression success and optimizing cell-free protein synthesis.
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