SYNTHETIC BZIP PROTEINS AS TARGETING AGENTS FOR PDT

以下NIH-AREA拨款提案描述了一项试点研究，旨在测试利用bZIP转录因子家族将光动力疗法（PDT）试剂靶向特定DNA序列（即AP-1元件：5'-TGAGTCA-3'）的功效。特别是，原生和从头设计的bZIP蛋白都将与光活性金属配合物衍生，这些配合物先前已被证明可以通过直接光诱导电子转移和/或单线态氧（1O2）的光化学产生引起DNA损伤。这些金属肽的体外DNA识别特性将通过电泳迁移转移试验进行研究，同时也将研究它们产生位点特异性DNA光损伤的能力。将对这些系统进行瞬态吸收和发射寿命研究，以帮助确定参与光蚀过程的机制。使用bZIP蛋白作为PDT试剂的靶向载体的一个优点是，它们的识别元件可以在大量的启动子中找到，这些启动子参与细胞分化和增殖等重要过程。这些部位的损伤应该具有很强的细胞毒性。提出的将PDT药物递送到特定细胞内靶点的方法的另一个优点是，原则上，它可以用于递送各种PDT药物。相信本提案中描述的工作将为鲍灵格林州立大学的学生提供一个新的机会，让他们参与一个多学科的研究项目，这可能会鼓励他们在生物医学科学领域进行研究生学习的兴趣。获得这一合同也将增强PI追求这一重点试点项目目标的能力。

The following proposal for a NIH-AREA grant describes a pilot study designed to test the efficacy of utilizing the family of bZIP transcription factors to target photodynamic therapy (PDT) reagents to specific sequences of DNA (i.e. the AP-1 element: 5'-TGAGTCA-3'). In particular, both native and de novo designed bZIP proteins will be derivatized with photoactive metal complexes which have been previously shown to cause DNA damage by either direct photoinduced electron-transfer and/or the photochemical generation of singlet oxygen (1O2). The in vitro DNA recognition properties of these metallopeptides will then be studied by electrophoretic mobility shift assays, as will be their ability to produce site-specific DNA photodamage. Transient absorption and emission lifetime studies will be conducted on these systems to help identify the mechanisms involved in the photolesion process. An advantage in using bZIP proteins as targeting vehicles for PDT reagents is that their recognition elements can be found within a large number of promoters which participate in such important processes as cell differentiation and proliferation. Damage to these sites should be very cytotoxic. An additional advantage in the proposed approach to deliver PDT drugs to specific intracellular targets is that it can, in principle, be used to deliver a wide variety of PDT drugs. It is believed that the work described in this proposal will provide a new opportunity for students at Bowling Green State University to participate in a multi-disciplinary research project which may encourage their interest in pursuing graduate studies in the biomedical sciences. Receipt of this award will also enhance the PI's ability to pursue the goals of this focused pilot-project.