Structural constraints on the display of foreign peptides on filamentous bacteriophages.
Strategies for the construction of vehicles for phage display are evaluated here on the basis of structural studies of filamentous bacteriophages. Potential sites for the insertion of foreign peptides into the major coat protein, gp8, of M13 are identified. Currently, the insertion of peptides into gp8 has two basic limitations: all insertion sites that have been used successfully are located within 5 amino acids (aa) of the N terminus, and in virions containing only mutant coat proteins, insertions larger than about 6 aa have not been successfully incorporated. The possible reasons for these limitations are discussed in terms of the structures of gp8 and the minor structural proteins, gp7 and gp9. Potential strategies for overcoming these limitations are outlined. Reasons for the successful incorporation of larger inserts into hybrid phage containing both native and mutant coat proteins are also discussed. The structures of gp6, gp7, and gp9 are described, and it is concluded that insertion sites in these minor proteins are unlikely to have substantial advantages over those currently being used in gp3. The structure of the coat protein of another filamentous phage, Pseudomonas phage Pf1, is also described. Its structure provides a number of clues for the successful design of phage display insertion sites. Because it contains a 7-aa surface loop in the major coat protein, the Pf1 coat protein may have significant advantages over gp8 of M13 as a vehicle for phage display.
Institute of Molecular Biophysics, Florida State University, Tallahassee 32306.
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Makowski
D017104:Bacteriophage M13 / Q000235:genetics / Q000648:ultrastructure*; D003001:Cloning, Molecular / Q000379:methods*; D003090:Coliphages / Q000235:genetics / Q000648:ultrastructure*; D004926:Escherichia coli / Q000235:genetics; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013499:Surface Properties; D014764:Viral Proteins / Q000648:ultrastructure*; D014771:Virion / Q000648:ultrastructure
D011506:Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins