HIV-1 NEF PROTEIN - PURIFICATION, CRYSTALLIZATIONS, AND PRELIMINARY-X-RAY DIFFRACTION STUDIES

Human immunodeficiency virus Nef protein accelerates virulent progress ion of AIDS by its interaction with specific cellular proteins involve d in cellular activation and signal transduction. Here we report the p urification and crystallization of the conserved core of HN-1(LAI) Nef protein in the unliganded form and in complex with the wild-type SH3 domain of the p59(fyn) protein-tyrosine kinase. One-dimensional NMR ex periments show that full-length protein and truncated fragment corresp onding to the product of HIV-1 protease cleavage have a well-folded co mpact tertiary structure. The ligand-free HIV 1 Nef(core) protein form s cubic crystals belonging to space group P23 with unit cell dimension s of a = b = c = 86.4 Angstrom. The Nef-Fyn SH3 cocrystals belong to t he space group P6(1)22 or its enantiomorph, P6(5)22, with unit cell di mensions of a = b = 108.2 Angstrom and c = 223.7 Angstrom. Both crysta l forms diffract to a resolution limit of 3.0 Angstrom resolution usin g synchrotron radiation, and are thus suitable for X-ray structure det ermination.