ArticleJournal of bacteriology1996
Topology of the membrane protein LamB by epitope tagging and a comparison with the X-ray model.
Article in Journal of bacteriology, 1996. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 40 citations in OpenAlex.
- Developing hybrid thiomaltodextrins as specific ligands to the maltodextrin transporter ofRSC advances · 2026Article
- Fluorescent Labeling of Outer Membrane Proteins Using the SpyCatcher-SpyTag System.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Components Subcellular Localization: Cell Surface Exposure.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Computational prediction of extracellular loops of the Por39 outer membrane porin ofComputational and structural biotechnology journal · 2023Article
- Outer membrane lipoprotein biogenesis: Lol is not the end.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2015Review
- Transmembrane domain of surface-exposed outer membrane lipoprotein RcsF is threaded through the lumen of β-barrel proteins.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Determination of surface-exposed, functional domains of gonococcal transferrin-binding protein A.Infection and immunity · 2004Article
- Site-directed mutagenesis of tyrosine 118 within the central constriction site of the LamB (maltoporin) channel of Escherichia coli. II. Effect on maltose and maltooligosaccharide binding kinetics.Biophysical journal · 2002Article
- Bacterial phage receptors, versatile tools for display of polypeptides on the cell surface.Journal of bacteriology · 2001Article
- Mutational analysis of the OprM outer membrane component of the MexA-MexB-OprM multidrug efflux system of Pseudomonas aeruginosa.Journal of bacteriology · 2001Article
- Engineering a tRNA and aminoacyl-tRNA synthetase for the site-specific incorporation of unnatural amino acids into proteins in vivo.Proceedings of the National Academy of Sciences of the United States of America · 1997Article
- Insertion mutagenesis of the lac repressor and its implications for structure-function analysis.Journal of bacteriology · 1997Article
- Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines.Nature biotechnology · 1997Review
- The levanase operon of Bacillus subtilis expressed in Escherichia coli can substitute for the mannose permease in mannose uptake and bacteriophage lambda infection.Journal of bacteriology · 1996Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We previously developed a genetic approach to study, with a single antibody, the topology of the outer membrane protein LamB, an Escherichia coli porin with specificity towards maltodextrins and a receptor for bacteriophage lambda. Our initial procedure consisted of inserting at random the same reporter epitope (the C3 neutralization epitope from poliovirus) into permissive sites of LamB (i.e., sites which tolerate insertions without deleterious effects on the protein activities or the cell). A specific monoclonal antibody was then used to examine the position of the inserted epitope with respect to the protein and the membrane. In the present work, we set up a site-directed procedure to insert the C3 epitope at new sites in order to distinguish between two-dimensional folding models. This allowed us to identify two new surface loops of LamB and to predict another periplasmic exposed region. The results obtained by random and directed epitope tagging are analyzed in light of the recently published X-ray structure of the LamB protein. Study of 23 hybrid LamB-C3 proteins led to the direct identification of five of the nine external loops (L4, L5, L6, L7, and L9) and led to the prediction of four periplasmic loops (I1, I4, I5, and I8) of LamB. Nine of the hybrid proteins did not lead to topological conclusions, and none led to the wrong predictions or conclusions. The comparison indicates that parts of models based on secondary structure predictions alone are not reliable and points to the importance of experimental data in the establishment of outer membrane protein topological models. The advantages and limitations of genetic foreign epitope insertion for the study of integral membrane proteins are discussed.
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