School of Pharmaceutical Science and Technology, Health Sciences Platform, Tianjin University, Tianjin 30072, China.
Pharmaceutical and Medicinal Chemistry (Computer-Aided Drug Design), Institute of Pharmacy, Freie Universität Berlin, Germany.
Biochem Pharmacol. 2020 Apr;174:113850. doi: 10.1016/j.bcp.2020.113850. Epub 2020 Feb 8.
The human cytochrome P450 enzyme CYP4Z1 remains an understudied enzyme despite its association with poor prognosis and overexpression in breast cancer. Hence, CYP4Z1 has previously been suggested as an anti-breast cancer target. In the present study we employed extended mutation analysis to increase our understanding of the substrate binding mode of this enzyme. In a combined in vitro and in silico approach we show for the first time that residue Arg487 plays an important role in substrate recognition and binding of CYP4Z1. Using a large array of recombinant CYP4Z1 mutants we show that, apart from Asn381, all other postulated binding residues only play an auxiliary role in substrate recognition and binding. Different substrate interaction motifs were identified via dynamic pharmacophores (dynophores) and their impact on catalytically competent substrate binding was classified. These new insights on the substrate recognition and binding mode represent an important step towards the rational design of CYP4Z1 prodrugs and guide further investigations into the so far poorly understood physiological role of CYP4Z1.
尽管人类细胞色素 P450 酶 CYP4Z1 与乳腺癌的预后不良和过表达有关，但它仍然是一种研究不足的酶。因此，CYP4Z1 以前被认为是一种抗乳腺癌的靶标。在本研究中，我们采用扩展的突变分析来增加对该酶底物结合模式的理解。通过体外和计算相结合的方法，我们首次表明残基 Arg487 在 CYP4Z1 的底物识别和结合中起重要作用。使用大量重组 CYP4Z1 突变体，我们表明，除了 Asn381 之外，所有其他假定的结合残基仅在底物识别和结合中起辅助作用。通过动态药效团（dynophores）鉴定了不同的底物相互作用基序，并对其对催化有效底物结合的影响进行了分类。这些关于底物识别和结合模式的新见解是朝着合理设计 CYP4Z1 前药迈出的重要一步，并指导对迄今为止了解甚少的 CYP4Z1 生理作用的进一步研究。
