Sequence-specific resonance assignments of the 1H-NMR spectra of a synthetic, biologically active EIAV Tat protein

Biochemistry. 1993 Aug 24;32(33):8439-45. doi: 10.1021/bi00084a008.


The equine infectious anemia virus (EIAV) trans-activating (Tat) protein is a close homologue of the human immunodeficiency virus (HIV) Tat protein. Both of these proteins bind to an RNA trans-activation responsive element (TAR). We synthesized chemically a protein with the sequence of the 75 amino acid Tat protein from EIAV. The chemically synthesized protein was shown to be biologically active. Circular dichroism (CD) and 1H nuclear magnetic resonance (NMR) spectroscopy were used to structurally characterize the synthetic protein. We obtained nearly complete resonance assignments in the 2D-NMR spectra of the protein at pH 3.0. There is at least some evidence from the experimental data that the basic TAR binding domain of the synthetic protein has a tendency to form a helix, but our experiments also indicate that the protein probably 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 trifluoroethanol/water solution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Circular Dichroism
  • Codon
  • Gene Products, tat / chemical synthesis
  • Gene Products, tat / chemistry*
  • Hydrogen
  • Infectious Anemia Virus, Equine / metabolism*
  • Magnetic Resonance Spectroscopy / methods
  • Molecular Sequence Data
  • Open Reading Frames
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Protein Structure, Secondary*


  • Codon
  • Gene Products, tat
  • Peptides
  • Hydrogen