Skip NavigationSkip to Content
Return Return to Search Results

A high-throughput fluorescence-anisotropy screen that identifies small molecule inhibitors of the DNA binding of B-ZIP transcription factors

  1. Author:
    Rishi, V.
    Potter, T.
    Laudeman, J.
    Reinhart, R.
    Silvers, T.
    Selby, M.
    Stevenson, T.
    Krosky, P.
    Stephen, A. G.
    Acharya, A.
    Moll, J.
    Oh, W. J.
    Scudiero, D.
    Shoemaker, R. H.
    Vinson, C.

  2. Author Address

    NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. SAIC, Frederick, MD 21702 USA. NCI, Dev Therapeut Program, Frederick, MD 21702 USA Vinson, C, NCI, Lab Metab, NIH, Bethesda, MD 20892 USA
    1. Year: 2005
    2. Date: MAY 15
  2. Journal: Analytical Biochemistry
    1. 340
    2. 2
    3. Pages: 259-271
  4. Type of Article: Article
  1. Abstract:

    We have developed a high-throughput fluorescence anisotropy screen, using a 384-well format, to identify small molecules that disrupt the DNA binding of B-ZIP proteins. Binding of a B-ZIP dimer to fluorescently labeled DNA can be monitored by fluorescence anisotropy. We screened the National Cancer Institute diversity, set of 1990 compounds to identify small molecules that disrupt the B-ZIP vertical bar DNA coniplex of CREB, C/EBP beta, VBP, and AP-1 (FOS vertical bar JUND) bound to their cognate DNA sequence. We identified 21 compounds that inhibited the DNA binding of at least one B-ZIP Protein, and 12 representative compounds were grouped depending on whether they displaced ethidium bromide from DNA. Of the 6 compounds that did not displace ethidium bromide. 2 also inhibited B-ZIP binding to DNA in a secondary electrophoretic niobility shift assay screen with some specificity. Thermal stability monitored by circular dichroism spectroscopy demonstrated that both compounds bound the basic region of the B-ZIP motif. NSC13778 preferentially binds C/EBP alpha of 1000-fold better than it binds C/EBP beta. Chimeric proteins combining C/EBP alpha. and C/EBP beta mapped the binding of NSC13778 to three amino acids immediately N terminal of the leucine zipper of C/EBP alpha.. These experiments suggest that the DNA binding of B-ZIP transcription factors is a potential target for clinical intervention. Published by Elsevier Inc

    See More

  1. Keywords:

External Sources

  1. View Full Text
  2. DOI: 10.1016/j.ab.2005.02.012
  3. View record in Web of Science®
  4. WOS: 000228779000010

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