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Site‐specific protein cleavage in vivo by an intein‐derived protease
Author(s) -
Volkmann Gerrit,
Volkmann Verena,
Liu Xiang-Qin
Publication year - 2012
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/j.febslet.2011.11.028
Subject(s) - intein , cleavage (geology) , protease , chemistry , in vivo , biochemistry , biology , enzyme , rna splicing , genetics , paleontology , rna , fracture (geology) , gene
Site‐specific protein cleavage is a ubiquitous process in cellular protein metabolism, yet molecular tools to provide control of protein cleavage inside living cells remain scarce. Here, we show that the C‐terminal intein fragment of the non‐canonical Ssp ( Synechocystis sp. PCC6803) DnaB S1 split‐intein can be used as a site‐specific protease for in vivo protein cleavage both in bacterial and eukaryotic cells. Mutagenesis data indicate a broad tolerance of the intein‐derived protease (IP) toward the amino acid upstream of the cleavage site. Furthermore, deletion studies reveal that the recognition sequence for the IP can be as short as ten amino acids. The structural features underlying the cleavage reaction preclude unintended proteolysis of endogenous proteins, thus ensuring that negative effects on cell viability are minimal.