Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24.
Mass spectrometry captures off-target drug binding and provides mechanistic insights into the human metalloprotease ZMPSTE24.
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疏水性药物的脱靶结合可通过特异性或非特异性结合非预期的膜蛋白靶点而导致不希望的副作用;然而，区分药物与膜蛋白的结合与去污剂、脂质和辅因子的结合是具有挑战性的。在这里，我们使用高分辨率质谱研究HIV蛋白酶抑制剂对人类锌金属蛋白酶ZMPSTE 24的影响。这种膜内蛋白酶在将前核纤层蛋白A转化为成熟核纤层蛋白A中起主要作用。我们通过ZMPSTE 24监测法尼基化的前核纤层蛋白A肽的蛋白水解，并且意外地发现C-末端肽产物与酶的保留。我们还解析了锌、脂质和HIV蛋白酶抑制剂的结合，并表明药物结合阻断了前层蛋白A肽切割并赋予ZMPSTE 24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早衰样副作用有关，而且还突出了记录脱靶药物结合的新方法。
Off-target binding of hydrophobic drugs can lead to unwanted side effects, either through specific or nonspecific 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 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.