Skip NavigationSkip to Content
Return Return to Search Results

Farnesylated and methylated KRAS4b: high yield production of protein suitable for biophysical studies of prenylated protein-lipid interactions

  1. Author:
    Gillette, W. K.
    Esposito, D.
    Abreu Blanco, M.
    Alexander, P.
    Bindu, L.
    Bittner, C.
    Chertov, O.
    Frank, P. H.
    Grose, C.
    Jones, J. E.
    Meng, Z.
    Perkins, S.
    Van, Q.
    Ghirlando, R.
    Fivash, M.
    Nissley, D. V.
    McCormick, F.
    Holderfield, M.
    Stephen, A. G.

  2. Author Address

    Cancer Research Technology Program, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc. PO Box B, Frederick, MD 21702. Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, 5 Memorial Drive, Bethesda MD 20892. Data Management Systems, NCI at Frederick, PO Box B, Frederick, MD 21702.
    1. Year: 2015
    2. Epub Date: 11/3/2015
  2. Journal: Scientific Reports
    1. 5
    2. Pages: 15916
  4. Type of Article: Article
  5. ISSN: 2045-2322 (Electronic) 2045-2322 (Linking)
  1. Abstract:

    Prenylated proteins play key roles in several human diseases including cancer, atherosclerosis and Alzheimer's disease. KRAS4b, which is frequently mutated in pancreatic, colon and lung cancers, is processed by farnesylation, proteolytic cleavage and carboxymethylation at the C-terminus. Plasma membrane localization of KRAS4b requires this processing as does KRAS4b-dependent RAF kinase activation. Previous attempts to produce modified KRAS have relied on protein engineering approaches or in vitro farnesylation of bacterially expressed KRAS protein. The proteins produced by these methods do not accurately replicate the mature KRAS protein found in mammalian cells and the protein yield is typically low. We describe a protocol that yields 5-10 mg/L highly purified, farnesylated, and methylated KRAS4b from insect cells. Farnesylated and methylated KRAS4b is fully active in hydrolyzing GTP, binds RAF-RBD on lipid Nanodiscs and interacts with the known farnesyl-binding protein PDEdelta.

    See More

  1. Keywords:

External Sources

  1. View Full Text
  2. DOI: 10.1038/srep15916
  3. PMID: 26522388
  4. PMCID: PMC4629113

Library Notes

  1. No notes added.
NCI at Frederick

You are leaving a government website.

This external link provides additional information that is consistent with the intended purpose of this site. The government cannot attest to the accuracy of a non-federal site.

Linking to a non-federal site does not constitute an endorsement by this institution or any of its employees of the sponsors or the information and products presented on the site. You will be subject to the destination site's privacy policy when you follow the link.

ContinueCancel