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Efficient introduction of alkene functionality into proteins in vivo
Author(s) -
van Hest Jan C.M,
Tirrell David A
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00489-x
Subject(s) - methionine , escherichia coli , dihydrofolate reductase , biochemistry , auxotrophy , chemistry , amino acid , in vivo , biology , enzyme , gene , microbiology and biotechnology
The methionine analogue 2‐amino‐5‐hexenoic acid (homoallylglycine, Hag) can be utilized by Escherichia coli in the initiation and elongation steps of protein biosynthesis. Use of an E. coli methionine auxotroph and Hag‐supplemented medium resulted in replacement of ca. 85% of the methionine residues in mouse dihydrofolate reductase expressed under control of a bacteriophage T5 promoter. N‐terminal sequencing indicated 92±5% occupancy of the initiator site by Hag. The vinyl function of Hag remains intact in the purified protein and suggests new chemistries for modification of natural and artificial proteins prepared in bacterial hosts.