Insertional mutagenesis in the tailspike protein of bacteriophage P22.
The tailspike protein (TSP) of bacteriophage P22 is a homotrimeric multifunctional protein responsible for recognition and hydrolysis of Salmonella typhimurium host receptors. Once properly folded, TSP shows an unusual stability to temperature and detergent denaturation, prompting the analysis of TSP as a framework for the positioning of heterologous protein segments. We have explored the flexibility of inner sites and both amino and carboxy termini to accommodate foreign peptides for phage display. In the examined inner sites, TSP is extremely sensitive to minor sequence modifications, the folding intermediates being rapidly degraded. However, both the amino and carboxy termini are tolerant to peptide fusions, rendering stable and functional chimeric proteins. Surprisingly, the amino terminus, which connects the tail to the neck structure, can accept large peptide fusions, and the foreign amino acid stretches are solvent-exposed and highly antigenic on assembled, infectious virus particles.
Carbonell
X
X
Institut de Biologia Fonamental, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Villaverde
A
A
D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics; D017100:Bacteriophage P22 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005814:Genes, Viral; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012486:Salmonella typhimurium / Q000821:virology; D012995:Solubility; D017376:Viral Tail Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology
D000956:Antigens, Viral; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D017376:Viral Tail Proteins; D006026:Glycoside Hydrolases; C083268:tailspike protein, bacteriophage