Wlodawer A
Macromolecular Structure Laboratory, NCI-Frederick Cancer Research and Development Center, Maryland 21702, USA.
Adv Virus Res. 1999;52:335-50. doi: 10.1016/s0065-3527(08)60305-x.
Crystal structures of the enzymatically competent catalytic domains of HIV-1 and ASV IN have been solved in the last few years. The structure of HIV-1 IN has been described only for apoenzyme and for a complex with Mg2+, whereas the structure of ASV IN has been presented as the apoenzyme, in the presence of divalent cations (Mn2+, Mg2+, Ca2+, Zn2+, and Cd2+), and with an inhibitor. A single ion of Mn2+, Mg2+, or Ca2+ interacts with the two aspartate side chains of the D,D(35)E catalytic center in octahedral coordination with four water molecules. However, two ions of Zn2+ or Cd2+ bind to the active site of IN with tetrahedral and octahedral coordination, respectively. Only small adjustments take place in the active site of ASV IN on binding of the metal cofactor(s), which are absolutely required for the activity of this enzyme. The placement of the side chains and metal ions in the active site is very similar to that observed even in distant members of this superfamily of polynucleotidyltransferases. Here the role of divalent cations in the enzymatic activity of IN and the search for inhibitors of this enzyme are discussed.
在过去几年中,已解析出HIV-1和ASV整合酶(IN)具有酶活性的催化结构域的晶体结构。HIV-1整合酶的结构仅针对无酶蛋白以及与Mg2+形成的复合物进行了描述,而ASV整合酶的结构则呈现为无酶蛋白、在存在二价阳离子(Mn2+、Mg2+、Ca2+、Zn2+和Cd2+)的情况下以及与一种抑制剂结合时的结构。单个Mn2+、Mg2+或Ca2+离子与D,D(35)E催化中心的两个天冬氨酸侧链相互作用,以八面体配位方式与四个水分子结合。然而,两个Zn2+或Cd2+离子分别以四面体和八面体配位方式结合到整合酶的活性位点。在结合金属辅因子时,ASV整合酶的活性位点仅发生微小调整,而金属辅因子是该酶活性绝对必需的。活性位点中侧链和金属离子的位置与在这个多聚核苷酸转移酶超家族的远亲成员中观察到的情况非常相似。本文讨论了二价阳离子在整合酶酶活性中的作用以及对该酶抑制剂的寻找。