Design and evolution of artificial M13 coat proteins

J Mol Biol. 2000 Jun 30;300(1):213-9. doi: 10.1006/jmbi.2000.3845.

Abstract

Using simple design and selective pressure, we have evolved an artificial M13 bacteriophage coat protein. M13 coat proteins first reside in the bacterial inner membrane and subsequently surround the DNA core of the assembled virus. The artificial coat protein (ACP) was designed and evolved to mimic both functions of the natural M13 coat proteins, but with an inverted orientation. ACP is a non-functional coat protein because it is not required for the production of phage particles. Instead, it incorporates into a phage coat which still requires all the natural coat proteins for structural integrity. In contrast with other M13 coat proteins, which can display polypeptides as amino-terminal fusions, ACP permits the carboxy-terminal display of large polypeptides. The results suggest that viruses can co-opt host membrane proteins to acquire new coat proteins and thus new functions. In particular, M13 bacteriophage can be engineered for new functions, such as carboxy-terminal phage display.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage M13* / chemistry
  • Bacteriophage M13* / genetics
  • Bacteriophage M13* / physiology
  • Capsid / chemistry*
  • Capsid / genetics*
  • Capsid / metabolism
  • Capsid Proteins*
  • Directed Molecular Evolution*
  • Enzyme-Linked Immunosorbent Assay
  • Membrane Proteins / chemistry*
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Molecular Sequence Data
  • Peptide Library
  • Protein Engineering*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Selection, Genetic
  • Sequence Alignment
  • Virus Assembly

Substances

  • Capsid Proteins
  • Membrane Proteins
  • Peptide Library
  • Recombinant Fusion Proteins
  • coat protein, Bacteriophage M13