HIV-1负调控因子蛋白不具有鸟嘌呤核苷酸结合、GTP酶或自身磷酸化活性。

The HIV-1 nef protein does not have guanine nucleotide binding, GTPase, or autophosphorylating activities.

作者信息

Backer J M, Mendola C E, Fairhurst J L, Kovesdi I

机构信息

Molecular Biology Research Section, American Cyanamid Company, Lederle Laboratories, Pearl River, NY 10965.

出版信息

AIDS Res Hum Retroviruses. 1991 Dec;7(12):1015-20. doi: 10.1089/aid.1991.7.1015.

DOI:10.1089/aid.1991.7.1015

PMID:1812942

Abstract

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.

摘要

已构建了具有苏氨酸 -15 或丙氨酸 -15 的重组 HIV-1 Nef 蛋白，并在 T7 细菌系统中表达。从细菌裂解物的可溶部分，Nef(苏氨酸 -15) 和 Nef(丙氨酸 -15) 在 MgCl2 存在下通过 6 步非变性色谱法纯化至接近均一性。两种纯化后的蛋白均未显示出先前报道的 GTP 结合活性。此外，Nef(苏氨酸 -15) 对 [γ-32P]GTP 或 [γ-32P]ATP 均无自磷酸化活性。虽然在 Nef 蛋白制剂中存在 GTP 酶活性，但在纯化过程中并未增加，且归因于细菌污染。