Open AccessSite-specific proteolysis of the Escherichia coli SecA protein in vivo
Author(s) -
Martin Mondigler,
Michael Ehrmann
Publication year - 1996
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.178.10.2986-2988.1996
Subject(s) - proteolysis , protease , biology , tobacco etch virus , cleavage (geology) , escherichia coli , biochemistry , amino acid , peptide sequence , enzyme , virus , virology , gene , plant virus , potyvirus , paleontology , fracture (geology)
A seven-amino-acid cleavage site specific for tobacco etch virus (TEV) protease was introduced into SecA at two separate positions after amino acids 195 and 252. Chromosomal wild-type secA was replaced by these secA constructs. Simultaneous expression of TEV protease led to cleavage of both SecA derivatives. In the functional SecA dimer, proteolysis directly indicated surface exposure of the TEV protease cleavage sites. Cleavage of SecA near residue 195 generated an unstable proteolysis product and a secretion defect, suggesting that this approach could be used to inactivate essential proteins in vivo.