A cysteine-free firefly luciferase retains luminescence activity.
A mutant of Photinus pyralis luciferase in which all four native cysteine residues are converted to serines retains about 10% of wild-type activity. This mutant should prove useful as a starting point for the introduction of biophysical probes of conformational changes associated with enzyme function. The activities of the cysteine-free mutant and others in which two or three cysteines are converted to serines suggest, however, that small chemical changes can have substantial and interdependent effects on bioluminescence. The introduction of probes should therefore be approached cautiously.
Kumita
J R
JR
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario, M5S 3H6, Canada.
Jain
L
L
Safroneeva
E
E
Woolley
G A
GA
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001517:Coleoptera / Q000201:enzymology*; D003545:Cysteine; D008156:Luciferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012694:Serine
D011994:Recombinant Proteins; D012694:Serine; D008156:Luciferases; D003545:Cysteine