Protein engineering with biosynthesized libraries from Bordetella bronchiseptica bacteriophage

PLoS One. 2013;8(2):e55617. doi: 10.1371/journal.pone.0055617. Epub 2013 Feb 7.

Abstract

Phage display offers a powerful approach to engineer protein affinity. A naturally occurring analog to phage display, the Bordetella bronchiseptica bacteriophage (BP) employs a highly variable protein termed the major tropism determinant (Mtd) to recognize its dynamic host. Propagation of BP provides a self-made phage library (SMPL) with vast numbers of phage particles, each displaying a single Mtd variant. We report applying the diversity of the BP-SMPL to access a tyrosine-rich library of Mtd variants. Expression of the SMPL-engineered Mtd variant as a GST-bound fusion protein demonstrated specific binding to the target T4 lysozyme with dissociation constants in the sub-micromolar range. The results guide future experiments with SMPLs applied to protein engineering.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Bacteriophages / metabolism*
  • Bordetella bronchiseptica / virology*
  • Molecular Sequence Data
  • Protein Engineering*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid

Substances

  • Recombinant Proteins