Loo J A, Holler T P, Sanchez J, Gogliotti R, Maloney L, Reily M D
Parke-Davis Pharmaceutical Research, Division of Warner Lambert Company, Ann Arbor, Michigan 48105, USA.
J Med Chem. 1996 Oct 11;39(21):4313-20. doi: 10.1021/jm960253w.
HIV nucleocapsid protein (NCp7) has been suggested as a possible target for 2,2'-dithiobis-[benzamide] and benzisothiazolone agents that inhibit viral replication in infected cells (Rice et al. Science 1995, 270, 1194-1197). The solution behavior of these compounds and the mechanistic events leading to removal of Zn from HIV nucleocapsid protein in vitro has been studied by electrospray ionization mass spectrometry, 500 MHz one- and two-dimensional nuclear magnetic resonance spectroscopy, and circular dichroism spectroscopy. We demonstrate that (1) Zn ejection is accompanied by formation of covalent complexes formed between the 2,2'-dithiobis[benzamide] monomers and Cys residues of Zn-depleted NCp7, (2) the rate of Zn ejection is faster for the C-terminal Zn finger and slower for the N-terminal finger, (3) Zn ejection results in a loss of structural integrity of the NCp7 protein, and (4) there is no appreciable interaction between a nonreactive isostere of the lead 2,2'-dithiobis[benzamide] and NCp7 in buffered aqueous solution. These findings are discussed in terms of the mechanism of action of Zn ejection by aromatic 2,2'-dithiobis[benzamides].
有人提出，HIV核衣壳蛋白（NCp7）可能是2,2'-二硫代双[苯甲酰胺]和苯并异噻唑啉酮类药物的作用靶点，这些药物可抑制感染细胞中的病毒复制（Rice等人，《科学》，1995年，第270卷，第1194 - 1197页）。通过电喷雾电离质谱、500兆赫的一维和二维核磁共振光谱以及圆二色光谱，研究了这些化合物的溶液行为以及在体外导致HIV核衣壳蛋白中锌去除的机制事件。我们证明：（1）锌的排出伴随着2,2'-二硫代双[苯甲酰胺]单体与锌耗尽的NCp7的半胱氨酸残基之间形成共价复合物；（2）C端锌指的锌排出速率较快，N端锌指的排出速率较慢；（3）锌的排出导致NCp7蛋白的结构完整性丧失；（4）在缓冲水溶液中，先导2,2'-二硫代双[苯甲酰胺]的非反应性等排体与NCp7之间没有明显的相互作用。根据芳香族2,2'-二硫代双[苯甲酰胺]排出锌的作用机制对这些发现进行了讨论。
