Open AccessA lipoprotein signal peptide plus a cysteine residue at the amino‐terminal end of the periplasmic protein β‐lactamase is sufficient for its lipid modification, processing and membrane localization in Escherichia coli
Author(s) -
Oudega Bauke,
Clark Dennis,
Stegehuis Freek,
Majoor Martijn J.,
Luirink Joen
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06127.x
Subject(s) - periplasmic space , cysteine , signal peptide , residue (chemistry) , biochemistry , chemistry , peptide , signal peptidase , bacterial outer membrane , membrane , peptide sequence , enzyme , escherichia coli , gene
By genetic exchange and in vitro mutagenesis a hybrid β‐lactamase was constructed that contained the pCloDF13‐encoded bacteriocin release protein signal peptide plus a cysteine residue coupled to the mature portion of β‐lactamase. Immunoblotting, labelling with [ 3 H]palmitate in the presence and absence of globomycin, and pulse‐chase experiments revealed that this hybrid construct is modified with lipid and processed into a lipid‐modified β‐lactamase. Subcellular localization studies revealed that this hybrid is localized both in the cytoplasmic and outer membranes of Escherichia coli cells. A mutant derivative with an incomplete lipobox (LVG instead of LVAC +1 ) was not processed and was found in the cytoplasmic membranes