title: The C-terminal domain of the HIV-1 Vif protein is natively unfolded in its unbound state.
abstract: The human immunodeficiency virus type-1 (HIV-1) Vif protein neutralizes the cellular defense mechanism against the virus. The C-terminal domain of Vif (CTD, residues 141-192) mediates many of its interactions. Full-length Vif is difficult to purify in large amounts, hence the only available structure of Vif is of residues 140-155 within the ElonginBC complex. Other structural information, derived from modeling and indirect experiments, indicates that the Vif CTD may be unstructured. Here, we chemically synthesized the Vif CTD using pseudo-proline-building blocks, studied its solution structure in the unbound state using biophysical techniques and found that it is unstructured under physiological conditions. The circular dichroism (CD) spectrum of Vif CTD showed a pattern of random coil with residual helical structure. The (15)N-HSQC nuclear magnetic resonance (NMR) spectrum was characteristic of natively unfolded peptides. Vif CTD eluted from an analytical gel filtration column earlier than expected, indicating an extended conformation. Disorder predictions found the CTD to be unstructured, in agreement with our experimental results. CD experiments showed that Vif CTD underwent a conformational change upon interacting with membrane-mimicking DPC micelles, but not upon binding to a peptide derived from its binding region in ElonginC. Our results provide direct evidence for the unfolded structure of the free Vif CTD and indicate that it may gain structure upon binding its natural ligands.
authors.affiliation: Institute of Chemistry, Hebrew University of Jerusalem, Safra Campus, Givat Ram, 91904 Jerusalem, Israel.
authors.forename: Tali H
authors.initials: TH
authors.lastname: Reingewertz
authors.forename: Hadar
authors.initials: H
authors.lastname: Benyamini
authors.forename: Mario
authors.initials: M
authors.lastname: Lebendiker
authors.forename: Deborah E
authors.initials: DE
authors.lastname: Shalev
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
mesh_terms: D001703:Biophysics; D002942:Circular Dichroism; D015497:HIV-1 / Q000235:genetics*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary / Q000235:genetics*; D054320:vif Gene Products, Human Immunodeficiency Virus / Q000235:genetics*
chemical_list: D054320:vif Gene Products, Human Immunodeficiency Virus; C514847:vif protein, Human immunodeficiency virus 1