title: A DsbA-Deficient Periplasm Enables Functional Display of a Protein with Redox-Sensitive Folding on M13 Phage.
abstract: The requirements for target protein folding in M13 phage display are largely underappreciated. Here we chose Fbs1, a carbohydrate binding protein, as a model to address this issue. Importantly, folding of Fbs1 is impaired in an oxidative environment. Fbs1 can be displayed on M13 phage using the SRP or Sec pathway. However, the displayed Fbs1 protein is properly folded only when Fbs1 is translocated via the SRP pathway and displayed using Escherichia coli cells with a DsbA-negative periplasm. This study indicates M13 phage display may be improved using a system specifically designed according to the folding requirements of each target protein.
authors.affiliation: New England Biolabs, Inc. , 240 County Road, Ipswich, Massachusetts 01938, United States.
authors.forename: Minyong
authors.initials: M
authors.lastname: Chen
authors.affiliation: New England Biolabs, Inc. , 240 County Road, Ipswich, Massachusetts 01938, United States.
authors.forename: James C
authors.initials: JC
authors.lastname: Samuelson
mesh_terms: D017104:Bacteriophage M13 / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D044783:F-Box Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D010084:Oxidation-Reduction; D019151:Peptide Library; D019897:Periplasm / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000172:deficiency*; D017510:Protein Folding; D055571:Secretory Pathway; D018271:Signal Recognition Particle
chemical_list: D018797:Cell Cycle Proteins; D029968:Escherichia coli Proteins; D044783:F-Box Proteins; C485508:FBXO2 protein, human; D009419:Nerve Tissue Proteins; D019151:Peptide Library; D018271:Signal Recognition Particle; D019704:Protein Disulfide-Isomerases; C526648:dsbA protein, E coli