Hou Yan, McGuinness Debra E, Prongay Andrew J, Feld Boris, Ingravallo Paul, Ogert Robert A, Lunn Charles A, Howe John A
Department of Virology, Schering-Plough Research Institute, Kenilworth, New Jersey 07033, USA.
J Biomol Screen. 2008 Jun;13(5):406-14. doi: 10.1177/1087057108317060. Epub 2008 May 14.
Small-molecule inhibitors of HIV integrase (HIV IN) have emerged as a promising new class of antivirals for the treatment of HIV/AIDS. The compounds currently approved or in clinical development specifically target HIV DNA integration and were identified using strand-transfer assays targeting the HIV IN/viral DNA complex. The authors have developed a second biochemical assay for identification of HIV integrase inhibitors, targeting the interaction between HIV IN and the cellular cofactor LEDGF/p75. They developed a luminescent proximity assay (AlphaScreen) designed to measure the association of the 80-amino-acid integrase binding domain of LEDGF/p75 with the 163-amino-acid catalytic core domain of HIV IN. This assay proved to be quite robust (with a Z' factor of 0.84 in screening libraries arrayed as orthogonal mixtures) and successfully identified several compounds specific for this protein-protein interaction.
HIV整合酶（HIV IN）的小分子抑制剂已成为治疗HIV/AIDS的一类有前景的新型抗病毒药物。目前已获批或处于临床开发阶段的化合物专门针对HIV DNA整合，并且是通过针对HIV IN/病毒DNA复合物的链转移测定法鉴定出来的。作者开发了第二种用于鉴定HIV整合酶抑制剂的生化测定法，该方法针对HIV IN与细胞辅因子LEDGF/p75之间的相互作用。他们开发了一种发光邻近测定法（AlphaScreen），旨在测量LEDGF/p75的80个氨基酸的整合酶结合结构域与HIV IN的163个氨基酸的催化核心结构域之间的结合。该测定法被证明相当可靠（在作为正交混合物排列的筛选文库中Z'因子为0.84），并成功鉴定出几种针对这种蛋白质-蛋白质相互作用的化合物。
