Bioorthogonal Approach to Identify Unsuspected Drug Targets in Live Cells

Authors

  • Dr. Katherine S. Yang,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Dr. Ghyslain Budin,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Dr. Carlos Tassa,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    Search for more papers by this author
  • Olivier Kister,

    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    Search for more papers by this author
  • Prof. Ralph Weissleder

    Corresponding author
    1. Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)
    2. Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115 (USA)
    • Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114 (USA)

    Search for more papers by this author

  • This work was supported by the National Institutes of Health (NIH) grant number RO1CA164448 and P50CA86355, K.Y. was supported by an NIH grant T32-CA79443.

Abstract

original image

A proteomics method to pull down secondary drug targets from live cells is described. The drug of interest is modified with trans-cyclooctene (TCO) and incubated with live cells. Upon cell lysis, the modified drug bound to the protein is pulled down using magnetic beads decorated with a cleavable tetrazine-modified linker. Samples are then run on an SDS-PAGE gel and isolated bands are submitted for mass spectrometry analysis to identify drug targets.

Ancillary