Conformational lability of herpesvirus protein VP22.
The herpesvirus protein VP22 traffics between cells, being exported from expressing cells in a non-Golgi-dependent manner and localizing in the nuclei of surrounding cells. This transport is retained in certain VP22 fusion proteins, making VP22 a candidate for use in macromolecular drug delivery. In an effort to understand the physical basis for this activity, we have initiated structural studies of VP22.C1, the C-terminal half of VP22, which possesses the full transport activity of the native protein. CD and Fourier transform infrared analyses indicate a secondary structure consisting of approximately 30% alpha-helix, 17% beta-sheet, and 51% disordered and turn structure. Unfolding studies conducted by CD, differential scanning calorimetry, and fluorescence reveal a series of discrete structural transitions in the range of 20-60 degrees C. CD and fluorescence studies of interactions between VP22.C1 and divalent cations and model polyanions indicate that Mg(2+), Zn(2+), oligonucleotides, and heparin interact with the protein, causing changes in secondary structure and thermal stability. Additionally, the interaction of VP22.C1 with model lipids was examined. Fluorescence titrations of the protein with trans-parinaric acid at various temperatures suggest the binding of one to two molecules of parinaric acid to VP22.C1 at temperatures >40 degrees C, suggesting the possibility of conformation dependent membrane interaction under physiological conditions.
Kueltzo
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LA
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas 66047, USA.
Normand
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N
O'Hare
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P
Middaugh
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CR
D002213:Capsid / Q000737:chemistry*; D002942:Circular Dichroism; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D016220:Fibroblast Growth Factor 1; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry; D005453:Fluorescence; D006493:Heparin / Q000494:pharmacology; D018259:Herpesvirus 1, Human / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D005231:Fatty Acids, Unsaturated; D016222:Fibroblast Growth Factor 2; D016220:Fibroblast Growth Factor 1; D006493:Heparin; C013641:parinaric acid