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Overexpression of recombinant proteins with a C‐terminal thiocarboxylate: Implications for protein semisynthesis and thiamin biosynthesis
Author(s) -
Kinsland Cynthia,
Taylor Sean V.,
Kelleher Neil L.,
Mclafferty Fred W.,
Begley Tadhg P.
Publication year - 1998
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.5560070821
Subject(s) - semisynthesis , intein , biosynthesis , biochemistry , chemistry , fusion protein , recombinant dna , cleavage (geology) , protein biosynthesis , biology , enzyme , gene , rna , rna splicing , paleontology , fracture (geology)
A facile and rapid method for the production of protein C‐terminal thiocarboxylates on DNA‐encoded polypeptides is described. This method, which relies on the mechanism of the cleavage reaction of intein‐containing fusion proteins, can produce multi‐milligram quantities of protein C‐terminal thiocarboxylate quickly and inexpensively. The utility of this method for protein semisynthesis and implications for studies on the biosynthesis of thiamin are discussed.