Selective destruction of protein function by chromophore-assisted laser inactivation.

TLDR

Protein function can be selectively inactivated by binding a malachite green–conjugated ligand or antibody and irradiating with a wavelength absorbed by the dye, sparing other cellular components. Chromophore‑assisted laser inactivation can be used to study cell‑surface phenomena by inactivating the functions of single proteins on living cells, a molecular extension of cellular laser ablation. The method employs a malachite green–conjugated ligand or antibody to bind the target protein, then a laser at the dye’s absorption wavelength selectively inactivates the bound protein while leaving other proteins unaffected. Chromophore‑assisted laser inactivation of protein function has been achieved, allowing ligand‑bound proteins in solution and on cell surfaces to be denatured without affecting other proteins, and it offers an advantage over genetics and inhibitors by enabling single‑cell inactivation via focused laser beams.

Abstract

Chromophore-assisted laser inactivation of protein function has been achieved. After a protein binds a specific ligand or antibody conjugated with malachite green (C.I. 42,000), it is selectively inactivated by laser irradiation at a wavelength of light absorbed by the dye but not significantly absorbed by cellular components. Ligand-bound proteins in solution and on the surfaces of cells can be denatured without other proteins in the same samples being affected. Chromophore-assisted laser inactivation can be used to study cell surface phenomena by inactivating the functions of single proteins on living cells, a molecular extension of cellular laser ablation. It has an advantage over genetics and the use of specific inhibitors in that the protein function of a single cell within the organism can be inactivated by focusing the laser beam.