期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 54, 期 29, 页码 8390-8394出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201501186
关键词
aptamers; biological activity; peptides; protein expression; synuclein
资金
- National Institutes of Health Research [R01-GM090323, R01-GM085006]
We report for the first time the recombinant expression of fully folded bioactive cyclotides inside live yeast cells by using intracellular protein trans-splicing in combination with a highly efficient split-intein. This approach was successfully used to produce the naturally occurring cyclotide MCoTI-I and the engineered bioactive cyclotide MCoCP4. Cyclotide MCoCP4 was shown to reduce the toxicity of human alpha-synuclein in live yeast cells. Cyclotide MCoCP4 was selected by phenotypic screening from cells transformed with a mixture of plasmids encoding MCoCP4 and inactive cyclotide MCoTI-I in a ratio of 1: 5 x 10(4). This demonstrates the potential for using yeast to perform phenotypic screening of genetically encoded cyclotide-based libraries in eukaryotic cells.
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