TY  - GEN
A1  - Hess, Anne-Katrin
A1  - Saffert, Paul
A1  - Liebeton, Klaus
A1  - Ignatova, Zoya
T1  - Optimization of translation profiles enhances protein expression and solubility
T2  - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe
N2  - mRNA is translated with a non-uniform speed that actively coordinates co-translational folding of protein domains. Using structure-based homology we identified the structural domains in epoxide hydrolases (EHs) and introduced slow-translating codons to delineate the translation of single domains. These changes in translation speed dramatically improved the solubility of two EHs of metagenomic origin in Escherichia coli. Conversely, the importance of transient attenuation for the folding, and consequently solubility, of EH was evidenced with a member of the EH family from Agrobacterium radiobacter, which partitions in the soluble fraction when expressed in E. coli. Synonymous substitutions of codons shaping the slow-transiting regions to fast-translating codons render this protein insoluble. Furthermore, we show that low protein yield can be enhanced by decreasing the free folding energy of the initial 5'-coding region, which can disrupt mRNA secondary structure and enhance ribosomal loading. This study provides direct experimental evidence that mRNA is not a mere messenger for translation of codons into amino acids but bears an additional layer of information for folding, solubility and expression level of the encoded protein. Furthermore, it provides a general frame on how to modulate and fine-tune gene expression of a target protein.
T3  - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 518 
KW  - transfer-RNA genes
KW  - codon usage
KW  - Escherichia coli
KW  - Epoxide hydrolases
KW  - messenger-RNA
KW  - sequence
KW  - elongation
KW  - Ribosome
KW  - mechanism
KW  - Membrane
Y1  - 2019
UR  - https://publishup.uni-potsdam.de/frontdoor/index/index/docId/40957
UR  - https://nbn-resolving.org/urn:nbn:de:kobv:517-opus4-409574
SN  - 1866-8372
IS  - 518
ER  -