ArticleThe Biochemical journal2004
A functional chimaeric S-layer-enhanced green fluorescent protein to follow the uptake of S-layer-coated liposomes into eukaryotic cells.
Article in The Biochemical journal, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 68 citations in OpenAlex.
- Bacterial nanotechnology as a paradigm in targeted cancer therapeutic delivery and immunotherapy.Microsystems & nanoengineering · 2024Review
- Slp-coated liposomes for drug delivery and biomedical applications: potential and challenges.International journal of nanomedicine · 2019Review
- S-Layer Protein-Based Biosensors.Biosensors · 2018Article
- Symmetry based assembly of a 2 dimensional protein lattice.PloS one · 2017Article
- S-layer fusion protein as a tool functionalizing emulsomes and CurcuEmulsomes for antibody binding and targeting.Colloids and surfaces. B, Biointerfaces · 2015Article
- Ion-specific control of the self-assembly dynamics of a nanostructured protein lattice.ACS nano · 2015Article
- Emulsomes meet S-layer proteins: an emerging targeted drug delivery system.Current pharmaceutical biotechnology · 2015Review
- S-layers: principles and applications.FEMS microbiology reviews · 2014Article
- Biomimetic interfaces based on S-layer proteins, lipid membranes and functional biomolecules.Journal of the Royal Society, Interface · 2014Review
- Production, secretion, and cell surface display of recombinant Sporosarcina ureae S-layer fusion proteins in Bacillus megaterium.Applied and environmental microbiology · 2012Article
- S-layer fusion proteins--construction principles and applications.Current opinion in biotechnology · 2011Review
- Composite S-layer lipid structures.Journal of structural biology · 2009Review
- Exploitation of the S-layer self-assembly system for site directed immobilization of enzymes demonstrated for an extremophilic laminarinase from Pyrococcus furiosus.Journal of biotechnology · 2008Article
- Structure, surface interactions, and compressibility of bacterial S-layers through scanning force microscopy and the surface force apparatus.Biophysical journal · 2006Article
- Expression and cytosolic assembly of the S-layer fusion protein mSbsC-EGFP in eukaryotic cells.Microbial cell factories · 2005Article
- Bacterial components as naturally inspired nano-carriers for drug/gene delivery and immunization: Set the bugs to work?Biotechnology advancesReview
Corrections and comments
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Authors and funding
9 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The chimaeric gene encoding a C-terminally truncated form of the S-layer protein SbpA of Bacillus sphaericus CCM 2177 and the EGFP (enhanced green fluorescent protein) was ligated into plasmid pET28a and cloned and expressed in Escherichia coli. Just 1 h after induction of expression an intense EGFP fluorescence was detected in the cytoplasm of the host cells. Expression at 28 degrees C instead of 37 degrees C resulted in clearly increased fluorescence intensity, indicating that the folding process of the EGFP moiety was temperature sensitive. To maintain the EGFP fluorescence, isolation of the fusion protein from the host cells had to be performed in the presence of reducing agents. SDS/PAGE analysis, immunoblotting and N-terminal sequencing of the isolated and purified fusion protein confirmed the presence of both the S-layer protein and the EGFP moiety. The fusion protein had maintained the ability to self-assemble in suspension and to recrystallize on peptidoglycan-containing sacculi or on positively charged liposomes, as well as to fluoresce. Comparison of fluorescence excitation and emission spectra of recombinant EGFP and rSbpA(31-1068)/EGFP revealed identical maxima at 488 and 507 nm respectively. The uptake of liposomes coated with a fluorescent monomolecular protein lattice of rSbpA(31-1068)/EGFP into HeLa cells was studied by confocal laser-scanning microscopy. The major part of the liposomes was internalized within 2 h of incubation and entered the HeLa cells by endocytosis.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.