The gag precursor contains a specific HIV‐1 protease cleavage site between the NC (P7) and P1 proteins

FEBS Letters

Wondrak, Ewald M.¹  Chermann, Jean-Claude¹  de Rocquigny, Hugues²  Louis, John M.³  Roques, Bernard P.² 

[1] Unité de Recherches sur les Retrovirus et Maladies Associées, INSERM U 322, Marseille, France;Université René Descartes (Paris V), UFR des Sciences Pharmaceutices et Biologiques, Paris, France;Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA

关键词: Human immunodeficiency virus;    HIV;    Nucleocapsid protein;    NC;    p7;    Protease;   

DOI  :  10.1016/0014-5793(93)80367-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The predicted protease cleavage site (p7/pl; [J. Virol. 66 (1992) 1856-1865]) within the nucleocapsid precursor protein (p15) of human immunodeficiency virus, type 1, was confirmed using an in vitro assay employing recombinant HIV-1 protease and a chemically synthesized 72 amino acid polypeptide containing the p7 and p1 protein domains of the native gag polyprotein. The cleavage occurred between amino acid 55 (N) and amino acid 56 (F) of the polypeptide, as determined by N-terminal sequencing. The hydrolysis was optimal at pH 6.0 and at high salt concentration. The kinetic parameters K _m, k _cat and K _cat/K _m were 99 μM (±8), 0.152 s⁻¹ (± 0.002) and 1.56 mM⁻¹· s⁻¹ (± 0.11), respectively. Reconstituted as well as denatured polypeptides were cleaved at approximately the same rate, demonstrating that the conformation of the p7 protein, as a result of the Zn²⁺-binding, had no significant effect on the rate of hydrolysis of the p7/pl cleavage.

【 授权许可】

Unknown