Light-regulated tetracycline binding to the Tet repressor.
Elucidation of the signal-transmission pathways between distant sites within proteins is of great importance in medical and bioengineering sciences. The use of optical methods to redesign protein functions is emerging as a general approach for the control of biological systems with high spatiotemporal precision. Here we report the detailed thermodynamic and kinetic characterization of novel chimeric light-regulated Tet repressor (TetR) switches in which light modulates the TetR function. Light absorbed by flavin mononucleotide (FMN) generates a signal that is transmitted to As-LOV and YtvA-LOV fused TetR proteins (LOV=light-oxygen-voltage), in which it alters the binding to tetracycline, the TetR ligand. The engineering of light-sensing protein modules with TetR is a valuable tool that deepens our understanding of the mechanism of signal transmission within proteins. In addition, the light-regulated changes of drug binding that we describe here suggest that engineered light-sensitive proteins may be used for the development of novel therapeutic strategies.
College of Pharmacy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 151-742 (Korea), Fax: (+82) 2-884-8334.
Jayoung
J
Moon
Jongsik
J
Gam
Seung-Goo
SG
Lee
Young-Ger
YG
Suh
Jeeyeon
J
Lee
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D005453:Fluorescence; D008027:Light; D008958:Models, Molecular; D010100:Oxygen; D017434:Protein Structure, Tertiary / Q000187:drug effects*; D013752:Tetracycline / Q000737:chemistry* / Q000494:pharmacology*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D013752:Tetracycline; D010100:Oxygen
DNA; LOV domain; antibiotics; nucleotides; proteins; tetracycline; thermodynamics