Site-specific proteolysis of the Escherichia coli SecA protein in vivo.
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.
Mondigler
M
M
Fakultät für Biologie, Universität Konstanz, Germany.
Ehrmann
M
M
D000251:Adenosine Triphosphatases / Q000502:physiology* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000502:physiology* / Q000648:ultrastructure; D001483:Base Sequence; D016384:Consensus Sequence; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000502:physiology* / Q000648:ultrastructure; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D010942:Plant Viruses / Q000201:enzymology; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000378:metabolism; D000069816:SEC Translocation Channels; D000081416:SecA Proteins; D013379:Substrate Specificity
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; D000069816:SEC Translocation Channels; D010450:Endopeptidases; C099488:TEV protease; D000251:Adenosine Triphosphatases; D000081416:SecA Proteins