Chromophore-assisted laser inactivation (CALI) to elucidate cellular mechanisms of cancer.
Chromophore-assisted laser inactivation (CALI) is a new technology for acute protein inactivation in living cells. It targets laser energy to specific proteins via non-function-blocking antibodies that are labeled with the dye malachite green. Excitation of the dye generates short-lived free radicals that damage the bound protein without affecting other cellular components. The wavelength of laser light used (620 nm) is not readily absorbed by cells such that non-specific light damage does not occur. CALI provides an alternative to other inactivation strategies and has the advantages of high spatial and temporal resolution. The ultimate value of this technology for cancer research will be assessed by how effective CALI is in ascribing in situ function during cancer-relevant processes and in identifying and validating protein targets for drug discovery. Recent work using CALI on ezrin and pp60-c-src, two proteins that may be involved in cancer, suggests its potential. Further application of CALI will likely be of utility for understanding cellular mechanisms of cancer and developing cancer therapeutics.
Jay
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DG
Department of Physiology, Tufts University School of Medicine, Boston, MA, USA. djay01@emerald.tufts.edu
Sakurai
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D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001426:Bacterial Proteins; D004396:Coloring Agents; D003598:Cytoskeletal Proteins / Q000737:chemistry; D029721:Drosophila Proteins; D018398:Homeodomain Proteins / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting; D007834:Lasers; D008565:Membrane Proteins / Q000737:chemistry; D018715:Microscopy, Video / Q000379:methods*; D009363:Neoplasm Proteins / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D010777:Photochemistry; D011506:Proteins / Q000737:chemistry* / Q000276:immunology; D012394:Rosaniline Dyes / Q000737:chemistry; D014157:Transcription Factors; D000099149:Ezrin
D000906:Antibodies; D001426:Bacterial Proteins; D004396:Coloring Agents; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D011506:Proteins; D012394:Rosaniline Dyes; D014157:Transcription Factors; D000099149:Ezrin; C052837:eve protein, Drosophila; C005095:malachite green