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PEDS Advance Access originally published online on July 25, 2009
Protein Engineering Design and Selection 2009 22(10):631-639; doi:10.1093/protein/gzp043
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© The Author 2009. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Mutations in fd phage major coat protein modulate affinity of the displayed peptide

G.A. Kuzmicheva1,4, P.K. Jayanna1, A.M. Eroshkin2, M.A. Grishina3, E.S. Pereyaslavskaya3, V.A. Potemkin3 and V.A. Petrenko1,5

1Department of Pathobiology, Auburn University, 252 Greene Hall, College of Veterinary Medicine, Auburn, AL 36849, USA 2 Burnham Institute for Medical Research, La Jolla, CA 92037, USA 3 Chelyabinsk State University, Chelyabinsk 454021, Russia

5 To whom correspondence should be addressed. E-mail: petreva{at}auburn.edu

Multibillion-clone libraries of phages displaying guest peptides fused to the major coat protein pVIII (landscape libraries) are a rich source of probes for proteinaceous and non-proteinaceous targets. As opposed to the pIII-type fusion phages, which display peptides as independent structural domains, the guest peptides in the pVIII-fusion phages can be structurally and functionally influenced by contiguous subunits. To decipher the impact of the locale of a guest peptide on its affinity characteristics, we constructed a library of phages carrying β-galactosidase-binding peptide ADTFAKSMQ at the N-terminus of the pVIII protein surrounded by random amino acids. It was found that mutagenesis of amino acids 12–19 (domain C) has polar effects on target binding affinity of the displayed peptide. The phages with highest affinity are characterized by: (i) a net electrostatic charge around –1 of domain C of the mutated phages at pH 7.0; (ii) a lower radius of cylinder coaxial to {alpha}-helix formed by domain C; (iii) a lower higher occupied molecular orbital (HOMO) of domain C leading to a decreased formation of hydrogen bonds and (iv) positively charged surface and torsion energy of domain C, which may require a conformational transition of N-terminal peptide ADTFAKSMQ for its binding with β-galactosidase. Influence of the guest peptide on the diversity of mutations in the neighboring landscape area was also observed.

Keywords: β-galactosidase/landscape phage/major coat protein pVIII/phage library

Received February 22, 2009; revised June 23, 2009; accepted June 24, 2009.

4 Present Address: Department of Chemistry, Sam Houston State University, 1003 Bowers Boulevard, r. CFS 239, Huntsville, TX 77340, USA


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