A DsbA-Deficient Periplasm Enables Functional Display of a Protein with Redox-Sensitive Folding on M13 Phage

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...

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Bibliographic Details
Published in:Biochemistry (Easton) 2016-06, Vol.55 (23), p.3175-3179
Main Authors: Chen, Minyong, Samuelson, James C
Format: Article
Language:English
Subjects:
Bacteriophage M13 - metabolism
Cell Cycle Proteins - chemistry
Cell Cycle Proteins - metabolism
Escherichia coli - metabolism
Escherichia coli Proteins
F-Box Proteins - chemistry
F-Box Proteins - metabolism
Humans
Nerve Tissue Proteins - chemistry
Nerve Tissue Proteins - metabolism
Oxidation-Reduction
Peptide Library
Periplasm - metabolism
Protein Disulfide-Isomerases - deficiency
Protein Folding
Secretory Pathway
Signal Recognition Particle
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Summary: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.
ISSN:0006-2960
1520-4995
DOI:10.1021/acs.biochem.6b00392