Premium
Interchange of aequorin and obelin bioluminescence color is determined by substitution of one active site residue of each photoprotein
Author(s) -
Stepanyuk Galina A.,
Golz Stefan,
Markova Svetlana V.,
Frank Ludmila A.,
Lee John,
Vysotski Eugene S.
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.01.004
Subject(s) - photoprotein , aequorin , bioluminescence , chemistry , stereochemistry , biochemistry , calcium , organic chemistry
The bioluminescence spectra from the Ca 2+ ‐regulated photoproteins aequorin ( λ max = 469 nm) and obelin ( λ max = 482 nm) differ because aequorin has an H‐bond from its Tyr82 to the bound coelenteramide, not present in obelin at the corresponding Phe88. Substitutions of this Phe88 by Tyr, Trp, or His shifted the obelin bioluminescence to shorter wavelength with F88Y having λ max = 453 nm. Removal of the H‐bond by the substitution of Y82F in aequorin shifted its bioluminescence to λ max = 501 nm. All mutants were stable with good activity and were expressible in mammalian cells, thereby demonstrating potential for monitoring multiple events in cells using multi‐color detection.