Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, UK.
Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK.
Nat Chem. 2016 Dec;8(12):1152-1158. doi: 10.1038/nchem.2591. Epub 2016 Aug 15.
Off-target binding of hydrophobic drugs can lead to unwanted side effects, either through specific or non-specific binding to unintended membrane protein targets. However, distinguishing the binding of drugs to membrane proteins from that of detergents, lipids and cofactors is challenging. Here, we use high-resolution mass spectrometry to study the effects of HIV protease inhibitors on the human zinc metalloprotease ZMPSTE24. This intramembrane protease plays a major role in converting prelamin A to mature lamin A. We monitored the proteolysis of farnesylated prelamin A peptide by ZMPSTE24 and unexpectedly found retention of the C-terminal peptide product with the enzyme. We also resolved binding of zinc, lipids and HIV protease inhibitors and showed that drug binding blocked prelamin A peptide cleavage and conferred stability to ZMPSTE24. Our results not only have relevance for the progeria-like side effects of certain HIV protease inhibitor drugs, but also highlight new approaches for documenting off-target drug binding.
疏水药物的非靶标结合可能导致不必要的副作用，其通过与非预期的膜蛋白靶标特异性或非特异性结合来实现。然而，区分药物与膜蛋白、洗涤剂、脂质和辅因子的结合具有挑战性。在这里，我们使用高分辨率质谱来研究 HIV 蛋白酶抑制剂对人类锌金属蛋白酶 ZMPSTE24 的影响。这种跨膜蛋白酶在将前层粘连蛋白 A 转化为成熟层粘连蛋白 A 中起主要作用。我们监测了 ZMPSTE24 对法尼基化前层粘连蛋白 A 肽的蛋白水解作用，出人意料地发现该酶与 C 端肽产物结合。我们还解析了锌、脂质和 HIV 蛋白酶抑制剂的结合，并表明药物结合阻止了前层粘连蛋白 A 肽的切割，并赋予 ZMPSTE24 稳定性。我们的研究结果不仅与某些 HIV 蛋白酶抑制剂药物的类早衰副作用有关，而且还突出了记录非靶标药物结合的新方法。
