A LIPOPROTEIN SIGNAL PEPTIDE PLUS A CYSTEINE RESIDUE AT THE AMINO-TERMINAL END OF THE PERIPLASMIC PROTEIN BETA-LACTAMASE IS SUFFICIENT FOR ITS LIPID MODIFICATION, PROCESSING AND MEMBRANE LOCALIZATION IN ESCHERICHIA-COLI
B. Oudega et al., A LIPOPROTEIN SIGNAL PEPTIDE PLUS A CYSTEINE RESIDUE AT THE AMINO-TERMINAL END OF THE PERIPLASMIC PROTEIN BETA-LACTAMASE IS SUFFICIENT FOR ITS LIPID MODIFICATION, PROCESSING AND MEMBRANE LOCALIZATION IN ESCHERICHIA-COLI, FEMS microbiology letters, 108(3), 1993, pp. 353-359
By genetic exchange and in vitro mutagenesis a hybrid beta-lactamase w
as constructed that contained the pCloDF13-encoded bacteriocin release
protein signal peptide plus a cysteine residue coupled to the mature
portion of beta-lactamase. Immunoblotting, labelling with [H-3]palmita
te in the presence and absence of globomycin, and pulse-chase experime
nts revealed that this hybrid construct is modified with lipid and pro
cessed into a lipid-modified beta-lactamase. Subcellular localization
studies revealed that this hybrid is localized both in the cytoplasmic
and outer membranes of Escherichia coli cells. A mutant derivative wi
th an incomplete lipobox (LVG instead of LVAC+1) was not processed and
was found in the cytoplasmic membranes