12th International Symposium on Bioluminescence & Chemiluminescence

Symposium abstracts:

Catalytic properties and bioluminescence spectra of recombinant firefly luciferases L. mingrelica with site mutation out of active site

Maloshenok, L.G.

Department of Chemistry, Lomonosov Moscow State University, 119899, Moscow, Russia

E-mail: mlg@enz.chem.msu.su

The influence of mutation of amino acid residues located out of active site of firefly luciferase L.mingrelica was studied. The positively charged residue His433 located near the surface of protein globule was chosen as the object of investigation. Mutants with substitutions His433® Asn, His433® Gln, His433® Ser have been made by the means of site-directed mutagenesis technique. The catalytic properties, spectra of bioluminescence, complexing of enzyme with substrates and their analogues were characterized. It was demonstrated that physico-chemical properties and structure of inserted amino acid residue provide noticeable influence on spectra of bioluminescence and change fluorescent properties of complexes of enzyme with luciferin and dimethyl luciferin. It was shown, that stability of luciferase mutants obtained is less in comparison with native recombinant firefly luciferase L.mingrelica.

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