The HIV-1 nef protein does not have guanine nucleotide binding, GTPase, or autophosphorylating activities.
Recombinant HIV-1 Nef proteins with either thr-15 or ala-15 have been constructed and expressed in the T7 bacterial system. From the soluble portion of bacterial lysates both Nef(thr-15) and Nef(ala-15) have been purified to near homogeneity through 6 nondenaturing chromatographic steps in the presence of MgCl2. Neither purified proteins display the previously reported GTP binding activity. Additionally Nef(thr-15) does not have autophosphorylating activity with either [gamma-32P]GTP or [gamma-32P]ATP. Although GTPase activity is present in the preparations of Nef proteins, it does not increase during purification and is attributed to bacterial contaminations.
Backer
J M
JM
Molecular Biology Research Section, American Cyanamid Company, Lederle Laboratories, Pearl River, NY 10965.
Mendola
C E
CE
Fairhurst
J L
JL
Kovesdi
I
I
D020558:GTP Phosphohydrolases / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D015693:Gene Products, nef / Q000302:isolation & purification / Q000378:metabolism*; D015754:Genes, nef; D006160:Guanosine Triphosphate / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics; D007700:Kinetics; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000378:metabolism; D054311:nef Gene Products, Human Immunodeficiency Virus
D015693:Gene Products, nef; D011994:Recombinant Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins