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Formation and Reversible Dissociation of Coiled Coil of Peptide to the C-Terminus of the HSV B5 Protein: A Time-Resolved Spectroscopic Analysis

Ordel J. Brown ^*, Santiago A. Lopez , A. Oveta Fuller  and Theodore Goodson, III ^*

^* Department of Chemistry, University of Michigan; and Department of Microbiology and Immunology, School of Medicine, University of Michigan, Ann Arbor, Michigan

Correspondence: Address reprint requests to Theodore Goodson III, Dept. of Chemistry, University of Michigan, Ann Arbor, MI 48109. E-mail: tgoodson{at}umich.edu.

An understanding of the molecular mechanisms of the newly characterized herpes simplex virus (HSV) B5 protein is important to further elucidate the HSV cell entry and infection. The synthetic peptide of B5 (wtB5) was functionalized with the nonlinear optical chromophore cascade yellow and its molecular dynamics was probed at physiological and endosomal pH (pH 7.4 and 5.5, respectively). Steady-state CD spectroscopy was utilized to characterize the peptides at different pH. These spectra showed structural changes in the peptide with time measured over several days. Nonlinear optical measurements were carried out to probe the interactions and local environment of the labeled peptide, and the increase in the two-photon cross section of this system suggests an increase in chromophore-peptide interactions. Time-resolved fluorescence upconversion measurements reflected changes in the hydrophilic and hydrophobic local environments of the labeled peptide-chromophore system. Ultrafast depolarization measurements gave rotational correlation times indicative of a reversible change in the size of the peptide. The time-resolved results provide compelling evidence of a reversible dissociation of the coiled coils of the wtB5 peptide. This process was found to be pH-insensitive. The data from this unique combination of techniques provide an initial step to understanding the molecular dynamics of B5 and a framework for the development of novel imaging methods based on two-photon emission, as well as new therapeutics for HSV.