A 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

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 [3H]palmitate in the presence and abse...

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Bibliographic Details
Published in:FEMS microbiology letters 1993-04, Vol.108 (3), p.353-349
Main Authors: Oudega, Bauke, Clark, Dennis, Stegehuis, Freek, Majoor, Martijn J., Luirink, Joen
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Anti-Bacterial Agents
Bacterial Outer Membrane Proteins - genetics
Bacterial Outer Membrane Proteins - metabolism
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacteriology
Base Sequence
beta-Lactamases - genetics
beta-Lactamases - metabolism
Biological and medical sciences
Biological Transport
Cysteine
Cytoplasm - metabolism
Endopeptidases - metabolism
Escherichia coli
Escherichia coli - metabolism
Escherichia coli Proteins
Fundamental and applied biological sciences. Psychology
Genetics
Lipid modification
Lipobox
Lipoproteins - metabolism
Membrane Proteins
Microbiology
Molecular Sequence Data
Palmitic Acid
Palmitic Acids - metabolism
Peptides - pharmacology
Processing
Protein Processing, Post-Translational - drug effects
Protein Sorting Signals - metabolism
Recombinant Fusion Proteins - metabolism
Serine Endopeptidases
Signal peptide
Subcellular localization
Substrate Specificity
β‐Lactamase
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Summary: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 [3H]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
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.1993.tb06127.x