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Incorporation of TePhe into Expressed Proteins is Minimally Perturbing
Author(s) -
Jia Bu Yong,
Nitz Mark
Publication year - 2021
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.202100160
Subject(s) - chemistry , amino acid , moiety , nuclear magnetic resonance spectroscopy , phenylalanine , side chain , biochemistry , stereochemistry , organic chemistry , polymer
Tellurium is a versatile heavy chalcogen with numerous applications in chemical biology, providing valuable probes in mass cytometry, fluorescence imaging and structural biology. L‐Tellurienylalanine (TePhe) is an analogue of the proteinogenic amino acid L‐phenylalanine (Phe) in which the phenyl side chain has been replaced by a 5‐membered tellurophene moiety. High incorporation level of TePhe in expressed proteins at defined sites is expected to facilitate studies in proteomics, protein NMR spectroscopy, and structure elucidation. As a model we chose immunoglobulin‐binding Protein G, B1 domain (GB1) to validate TePhe as a suitable structural analogue for Phe. We demonstrate that approximately 1 in 2 of all Phe sites within GB1 can be substituted with TePhe through expression in standard non‐Phe‐auxotrophic E. coli in Phe‐deficient media containing glyphosate, an inhibitor of aromatic amino acid biosynthesis. The TePhe content of the GB1 sample can be further increased to 85 % through HPLC. Using NMR and CD spectroscopy, we confirm that the Phe‐to‐TePhe substitution has negligible impact on the global structure and stability of GB1.