A general system for evaluating the efficiency of chromophore-assisted light inactivation (CALI) of proteins reveals Ru(II) tris-bipyridyl as an unusually efficient "warhead".
Chromophore-assisted light inactivation (CALI) of proteins is a potentially powerful tool in biological research for the triggered disruption of protein function. It involves the creation of chimeric molecules that can bind specifically to the protein target and can also sensitize the photo-generation of singlet oxygen, which inactivates the target protein. There remains a need for more efficient chromophores for singlet oxygen generation. Here we report a general and convenient system with which to evaluate the efficiency of chromophores in CALI both in crude extracts and in living cells. We employ this system to show that a readily available derivative of ruthenium(II) tris-bipyridyl dication is an unusually efficient "warhead" for CALI, exhibiting a performance markedly superior to the commonly used organic fluorophore, fluorescein.
Lee
Jiyong
J
Division of Translational Research, Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390-9185, USA.
Yu
Peng
P
Xiao
Xiangshu
X
Kodadek
Thomas
T
D002384:Catalysis; D016503:Drug Delivery Systems; D019793:Fluorescein / Q000737:chemistry / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008027:Light; D008954:Models, Biological; D009942:Organometallic Compounds / Q000494:pharmacology*; D010789:Phototherapy / Q000379:methods*; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology; D012428:Ruthenium / Q000737:chemistry / Q000494:pharmacology*; D026082:Singlet Oxygen / Q000494:pharmacology
D005456:Fluorescent Dyes; D009942:Organometallic Compounds; D011506:Proteins; D011993:Recombinant Fusion Proteins; D026082:Singlet Oxygen; D012428:Ruthenium; D019793:Fluorescein