Prolipoprotein signal peptidase of Escherichia coli requires a cysteine residue at the cleavage site.
A signal peptidase specifically required for the secretion of the lipoprotein of the Escherichia coli outer membrane cleaves off the signal peptide at the bond between a glycine and a cysteine residue. This cysteine residue was altered to a glycine residue by guided site-specific mutagenesis using a synthetic oligonucleotide and a plasmid carrying an inducible lipoprotein gene. The induction of mutant lipoprotein production was lethal to the cells. A large amount of the prolipoprotein was accumulated in the outer membrane fraction. No protein of the size of the mature lipoprotein was detected. These results indicate that the prolipoprotein signal peptidase requires a glyceride modified cysteine residue at the cleavage site.
Inouye
S
S
Department of Biochemistry, State University of New York at Stony Brook, 11794, USA.
Franceschini
T
T
Sato
M
M
Itakura
K
K
Inouye
M
M
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D029968:Escherichia coli Proteins; D015870:Gene Expression; D005798:Genes, Bacterial; D008074:Lipoproteins; D008565:Membrane Proteins; D016296:Mutagenesis; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*
D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D008074:Lipoproteins; C099932:Lpp protein, E coli; D008565:Membrane Proteins; D011498:Protein Precursors; D012697:Serine Endopeptidases; C026515:type I signal peptidase; D003545:Cysteine