12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Light-induced fluorescence kindling in GFP-like chromoproteins can be
switched on/off and modified by mutagenesis

Chudakov, Dmitry M., Lukyanov, Konstantin A., Lukyanov, Sergey A.

Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117997 Moscow, Russia

Email: luk@ibch.ru

GFP-like chromoprotein from Anemonia sulcata asFP595 (asCP) is able to become red fluorescent (to "kindle") in response to intensive green light irradiation. After kindling this protein relax to the initial non-fluorescent state for about 1-2 minutes, but can be brought to the initial state (to be bleached) instantly by short blue light irradiation. Both kindling and bleaching are reversible processes. Here we used direct and random mutagenesis to localize amino acid positions crucial for this photoconversion. Several positions were found to play a key role in control of kindling/bleaching behavior. Some mutants completely lost the photoconversion ability, while others possessed
significantly changed relaxation kinetics. We applied key amino acid substitutions to the two other chromoproteins, hcCP and cgCP, derived from Heteractis crispa and Condylactis gigantea species. These substitutions resulted in appearance of kindling and bleaching properties in these chromoproteins. A model of molecular events underlying the photoconversion in chromoproteins is proposed.

This is a preprint of an article accepted for publication in Luminescence: Copyright 2001 John
Wiley & Sons, Ltd (Wiley website)