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Nucleotide Exchange on RAS Proteins Using Hydrolysable and Non-hydrolysable Nucleotides

  1. Author:
    Waybright,Tim
    Stephen,Andy

  2. Author Address

    NCI RAS Initiative, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Frederick, MD, USA. Timothy.Waybright@nih.gov.,
    1. Year: 2024
  2. Journal: Methods in Molecular Biology (Clifton, N.J.)
    1. 2797
    2. Pages: 35-46
  4. Type of Article: Article
  1. Abstract:

    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 GDPßS, 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. © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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External Sources

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  2. DOI: 10.1007/978-1-0716-3822-4_4
  3. PMID: 38570451

Library Notes

  1. Fiscal Year: FY2023-2024
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