Hydrophobicity engineering to facilitate surface display of heterologous gene products on Staphylococcus xylosus. 1995

T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
Centre d'Immunologie Pierre Fabre, Saint-Julien en Genevois, France.

Protein engineering has been employed to investigate the effect of specific amino acid changes on the targeting of heterologous proteins to the outer cell surface of the Gram-positive bacterium Staphylococcus xylosus. Three different variants, corresponding to a 101 amino acid region of the major glycoprotein (G protein) of human respiratory syncytial virus (RSV), were generated in which multiple hydrophobic phenylalanine residues were either substituted or deleted. The different G protein fragments were expressed as one part of recombinant receptors designed for surface display on S. xylosus cells. The engineered variants of the RSV G protein hybrid receptors were, in contrast to a non-engineered fragment, efficiently targeted to the outer cell surface of recombinant S. xylosus cells as determined by different methods, including fluorescence-activated cell sorting. In addition, immunization of mice with live recombinant S. xylosus demonstrated that surface exposure was required to generate receptor-specific antibodies. The present strategy of hydrophobic engineering should be of general interest in surface-display applications and for secretion of proteins otherwise difficult to translocate through host cell membranes.

UI MeSH Term Description Entries
D008565 Membrane Proteins Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors. Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D008807 Mice, Inbred BALB C An inbred strain of mouse that is widely used in IMMUNOLOGY studies and cancer research. BALB C Mice, Inbred,BALB C Mouse, Inbred,Inbred BALB C Mice,Inbred BALB C Mouse,Mice, BALB C,Mouse, BALB C,Mouse, Inbred BALB C,BALB C Mice,BALB C Mouse
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
D012136 Respiratory Syncytial Viruses A group of viruses in the PNEUMOVIRUS genus causing respiratory infections in various mammals. Humans and cattle are most affected but infections in goats and sheep have also been reported. Chimpanzee Coryza Agent,Orthopneumovirus,RSV Respiratory Syncytial Virus,Chimpanzee Coryza Agents,Coryza Agent, Chimpanzee,Orthopneumoviruses,Respiratory Syncytial Virus,Syncytial Virus, Respiratory,Virus, Respiratory Syncytial
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D003124 Colorimetry Any technique by which an unknown color is evaluated in terms of standard colors. The technique may be visual, photoelectric, or indirect by means of spectrophotometry. It is used in chemistry and physics. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
D005260 Female Females
D005434 Flow Cytometry Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake. Cytofluorometry, Flow,Cytometry, Flow,Flow Microfluorimetry,Fluorescence-Activated Cell Sorting,Microfluorometry, Flow,Cell Sorting, Fluorescence-Activated,Cell Sortings, Fluorescence-Activated,Cytofluorometries, Flow,Cytometries, Flow,Flow Cytofluorometries,Flow Cytofluorometry,Flow Cytometries,Flow Microfluorometries,Flow Microfluorometry,Fluorescence Activated Cell Sorting,Fluorescence-Activated Cell Sortings,Microfluorimetry, Flow,Microfluorometries, Flow,Sorting, Fluorescence-Activated Cell,Sortings, Fluorescence-Activated Cell
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle

Related Publications

T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
June 1993, Gene,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
July 1997, Applied and environmental microbiology,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
January 2019, Frontiers in microbiology,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
January 1990, Methods in enzymology,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
May 2008, Research in microbiology,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
June 1997, FEMS microbiology letters,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
February 2010, Molecular biotechnology,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
August 2006, Meat science,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
January 1992, Research in veterinary science,
T N Nguyen, and M H Gourdon, and M Hansson, and A Robert, and P Samuelson, and C Libon, and C Andréoni, and P A Nygren, and H Binz, and M Uhlén
February 1993, Journal of bacteriology,
Copied contents to your clipboard!