使用可水解核苷酸和不可水解核苷酸进行 RAS 蛋白的核苷酸交换。
Nucleotide Exchange on RAS Proteins Using Hydrolysable and Non-hydrolysable Nucleotides.
机构信息
NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA.
出版信息
Methods Mol Biol. 2024;2797:35-46. doi: 10.1007/978-1-0716-3822-4_4.
Biochemical and biophysical assays using recombinant RAS require the protein to be in either the active or inactive state. Here we describe methods to exchange the nucleotide present in the purified RAS protein with either GDPβS, GppNHp, or GTP depending on the assay requirement. In addition, we also describe the HPLC method used to validate the exchange process and provide information on the efficiency of the nucleotide exchange.
使用重组 RAS 进行生化和生物物理测定需要蛋白质处于激活或失活状态。在这里，我们描述了根据测定要求将存在于纯化 RAS 蛋白中的核苷酸与 GDPβS、GppNHp 或 GTP 交换的方法。此外，我们还描述了用于验证交换过程的 HPLC 方法，并提供有关核苷酸交换效率的信息。
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