Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 109; no. 12; pp. E690 - E697
• Main Authors: Zakeri, Bijan, Fierer, Jacob O, Celik, Emrah, Chittock, Emily C, Schwarz-Linek, Ulrich, Moy, Vincent T, Howarth, Mark
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 20.03.2012; National Acad Sciences
• Series: PNAS Plus
• Subjects: Adhesins; Adhesins, Bacterial - metabolism; amides; Amides - chemistry; Bacteria; Biological Sciences; Biophysics - methods; Boiling; Cell Membrane - metabolism; Cell surface; Cells; Chemical bonds; engineering; Fibronectin-binding protein; Fibronectins - chemistry; HeLa Cells; Humans; Hydrogen-Ion Concentration; Mammalian cells; mammals; Microscopy, Atomic Force - methods; mixing; Molecular Sequence Data; Molecules; Peptides; Peptides - chemistry; pH effects; PNAS Plus; Protein Binding; Protein Engineering - methods; Protein interaction; Protein Structure, Tertiary; Proteins; Spectrometry, Mass, Electrospray Ionization - methods; Spectroscopy; Spectrum analysis; Splitting; Streptococcus pyogenes; Streptococcus pyogenes - metabolism; Temperature; Temperature effects; Thermodynamics

Protein interactions with peptides generally have low thermodynamic and mechanical stability. Streptococcus pyogenes fibronectin-binding protein FbaB contains a domain with a spontaneous isopeptide bond between Lys and Asp. By splitting this domain and rational engineering of the fragments, we obtai...