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The Basic Structure of Filamentous Phage and Its Use in the Display of Combinatorial Peptide Libraries

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Combinatorial Peptide Library Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 87))

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Abstract

The filamentous phage constitutes a large number of male-specific bacteriophage having similar shape, size, and lifecycle. The most studied phage are f1,fd,and M13, all of which Infect E colz cells through their F pill. This group of F-spectfic phage share 98% homology in their DNA sequence. Their nme coding genes (coding for ten proteins), as well as then two mtergenic regions, are ordered and oriented in the same way. The phage particle has a filamentous shape of 930-nm in length, a diameter of 6.5-nm, and contains a ssDNA of about 6400 bp, packaged within five coat proteins named pVII1, p111, pV1, pVI1, and pIX ( Fig. 1 ). The major coat protein, pVII1, wraps the phage DNA with 2800 of its copies arranged in a helical symmetry (1). When the phage DNA is elongated as a result of DNA insertion, the number of pVII1 copies mcreases to compensate for the increased length. (2,3). At the proximal end of the virion (the first to cross the membrane when the phage leaves the host cell), five copies of pVII and of pIX form a 30Å, plug structure. These are two small hydrophobic peptides, 33 and 32 kDa, respectively, that play a role in the early stages of phage assembly, where they serve as a nucleus for the subsequent deposition of pVII1 (Fig. 2) In their absence, almost no phage particles are formed (4,5). The distal end of the virion terminates with five copies of pIII and of pVI (6–8). These protems are required for terminating the deposition of pVIII during phage assembly and for anchoring the phage to the bacterium pili (5). Negative staining of phage particles shows that pII1 and pVI form a cylindrical shape with a pointed end, and pIII extends further as a thin protrusion (9).

A “fish scale” structure of filamentous phage. Symbols: O-library insertion sites; gray- hydrophobic surface; white- hydrophilic surface.

Assembly of the phage virion and its release from E. coli cells.

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Authors and Affiliations

  1. Department of Membrane Research and Biophysics, The Weizmann Institute of Science, Rehovot, Israel

    Shmuel Cabilly

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  1. Shmuel Cabilly
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  1. Weizmann Institute of Science, Rehovot, Israel

    Shmuel Cabilly

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© 1998 Humana Press Inc., Totowa, NJ

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Cabilly, S. (1998). The Basic Structure of Filamentous Phage and Its Use in the Display of Combinatorial Peptide Libraries. In: Cabilly, S. (eds) Combinatorial Peptide Library Protocols. Methods in Molecular Biology™, vol 87. Humana Press. https://doi.org/10.1385/0-89603-392-9:129

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  • DOI: https://doi.org/10.1385/0-89603-392-9:129

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-392-4

  • Online ISBN: 978-1-59259-571-6

  • eBook Packages: Springer Protocols

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