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Cotranslational Incorporation of a Structurally Diverse Series of Proline Analogues in an Escherichia coli Expression System
Author(s) -
Kim Wookhyun,
George Anna,
Evans Melissa,
Conticello Vincent P.
Publication year - 2004
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200400052
Subject(s) - escherichia coli , proline , amino acid , biochemistry , context (archaeology) , biology , substrate (aquarium) , function (biology) , recombinant dna , chemistry , gene , genetics , paleontology , ecology
A set of Escherichia coli expression strains have been defined that are competent for the incorporation of a structurally diverse series of proline analogues under culture conditions that are compatible with high levels of analogue substitution within a proline‐rich protein substrate. These bacterial strains have been employed to assay the efficacy of incorporation of noncanonical amino acids into a recombinant‐protein test substrate and to create variant polypeptides in which native protein sequences have been globally substituted with imino acid analogues in response to proline codons. We envision that these methods may be used to interrogate the effect of imino acid substitution on protein structure and function and may be particularly informative in the context of structural comparison of a series of modified proteins with respect to the stereoelectronic differences between the incorporated proline analogues.