SEQUENCE-SPECIFIC RESONANCE ASSIGNMENTS OF THE H-1-NMR SPECTRA OF A SYNTHETIC, BIOLOGICALLY-ACTIVE EIAV TAT PROTEIN

The equine infectious anemia virus (EIAV) trans-activating (Tat) prote in is a close homologue of the human immunodeficiency virus (HIV) Tat protein. Both of these proteins bind to an RNA trans-activation respon sive element (TAR). We synthesized chemically a protein with the seque nce of the 75 amino acid Tat protein from EIAV. The chemically synthes ized protein was shown to be biologically active. Circular dichroism ( CD) and H-1 nuclear magnetic resonance (NMR) spectroscopy were used to structurally characterize the synthetic protein. We obtained nearly c omplete resonance assignments in the 2D-NMR spectra of the protein at PH 3.0. There is at least some evidence from the experimental data tha t the basic TAR binding domain of the synthetic protein has a tendency to form a helix, but our experiments also indicate that the protein p robably does not have an overall stable tertiary structure in aqueous solution at this pH. CD spectroscopy suggested that the protein adopts a more stable, predominantly alpha-helical structure in a trifluoroet hanol/water solution.