HIV-1 Nef protein: purification, crystallizations, and preliminary X-ray diffraction studies.
Human immunodeficiency virus Nef protein accelerates virulent progression of AIDS by its interaction with specific cellular proteins involved in cellular activation and signal transduction. Here we report the purification and crystallization of the conserved core of HIV-1LAI Nef protein in the unliganded form and in complex with the wild-type SH3 domain of the P59fyn protein-tyrosine kinase. One-dimensional NMR experiments show that full-length protein and truncated fragment corresponding to the product of HIV-1 protease cleavage have a well-folded compact tertiary structure. The ligand-free HIV-1 Nefcore protein forms cubic crystals belonging to space group P23 with unit cell dimensions of a = b = c = 86.4 A. The Nef-Fyn SH3 cocrystals belong to the space group P6(1)22 or its enantiomorph, P6(5)22, with unit cell dimensions of a = b = 108.2 A and c = 223.7 A. Both crystal forms diffract to a resolution limit of 3.0 A resolution using synchrotron radiation, and are thus suitable for X-ray structure determination.
Franken
P
P
Centre de Biochimie Structurale, UMR C9955 CNRS, U414 INSERM, Université Montpellier I, Faculté de Pharmacie, France.
Arold
S
S
Padilla
A
A
Bodeus
M
M
Hoh
F
F
Strub
M P
MP
Boyer
M
M
Jullien
M
M
Benarous
R
R
Dumas
C
C
D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D015693:Gene Products, nef / Q000737:chemistry* / Q000302:isolation & purification; D016333:HIV Protease / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry*; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D010446:Peptide Fragments / Q000737:chemistry; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D010446:Peptide Fragments; D011994:Recombinant Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D016333:HIV Protease