SBS: Mgr. SBS: Jan Novotný, Ph.D.: Elucidation of (supra)molecular structure by using paramagnetic effects in NMR

The development of light-emitting luciferase enzymes for ultra-sensitive biological tests is often only possible if the basic catalytic mechanism is thoroughly understood. Therefore, knowledge of the Michaelis enzyme-substrate complex at the atomic level, which reveals the molecular details of luciferin recognition and catalytic chemistry, is essential for understanding and subsequently rationally improving bioluminescent reactions. However, many known luciferases exploit the enormous conformational space of proteins, which often prevents complete structural characterization. In addition, the use of related luciferins is problematic because their conversion to oxyluciferin in the presence of luciferase prevents the capture of the enzyme-luciferin conformation in the activated state. In this lecture, I will outline how to deal with such obstacles and focus on recent discoveries of marine bioluminescence mechanisms, which have been made possible by combining a modified primordial enzyme and the availability of a non-oxidizable luciferin analog.