The role of active site residue arginine 218 in firefly luciferase bioluminescence

Biochemistry
B R BranchiniNathan C Portier

Abstract

Firefly luciferase catalyzes the highly efficient emission of yellow-green light from substrate firefly luciferin by a sequence of reactions that require Mg-ATP and molecular oxygen. We had previously developed a working model of the luciferase active site based on the X-ray structure of the enzyme without bound substrates. In our model, the side chain guanidinium group of Arg218 appears to be located in close proximity to the substrate's hydroxyl group at the bottom of the luciferin binding pocket. A similar role for Arg337 also has been proposed. We report here the construction, purification, and characterization of mutant luciferases R218A, R218Q, R218K, R337Q, and R337K. Alteration of the Arg218 side chain produced enzymes with 15-20-fold increases in the Km values for luciferin. The contrasting near-normal Km values for luciferin determined with the Arg337 enzymes support our proposal that Arg218 (and not Arg337) is an essential luciferin binding site residue. Bioluminescence emission studies indicated that in the absence of a positively charged group at position 218, red bioluminescence was produced. Based on this result and those of additional fluorescence experiments, we speculate that Arg218 maintains the polarity and ...Continue Reading

References

Aug 1, 1997·The Journal of Biological Chemistry·B R BranchiniM H Murtiashaw
Jan 19, 1999·Annual Review of Cell and Developmental Biology·T Wilson, J W Hastings

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Citations

May 15, 2002·The International Journal of Biochemistry & Cell Biology·Xi Cheng WangHai Meng Zhou
Apr 11, 2009·Journal of the American Chemical Society·Fengyi LiuRoland Lindh
Jan 23, 2009·Journal of the American Chemical Society·Takashi HiranoDaisuke Hashizume
Sep 3, 2009·Journal of the American Chemical Society·Pance NaumovShunichi Fukuzumi
Feb 12, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·V R VivianiY Ohmiya
Feb 12, 2008·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Hugo Fraga
Dec 22, 2009·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·A J Silva NetoV R Viviani
Oct 5, 2010·Physical Chemistry Chemical Physics : PCCP·Bruce F MilneFernando Nogueira
Jun 8, 2010·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Vadim R VivianiJoão A R G Barbosa
Feb 16, 2011·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Mikhail I Koksharov, Natalia N Ugarova
Apr 21, 2011·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·R A PradoV R Viviani
Sep 24, 2013·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Mikhail I Koksharov, Natalia N Ugarova
Aug 8, 2009·Protein Engineering, Design & Selection : PEDS·Ali Riahi-Madvar, Saman Hosseinkhani
Oct 1, 2004·Molecular and Cellular Biology·Magdalena Rakwalska, Sabine Rospert
Oct 28, 2009·Cellular and Molecular Life Sciences : CMLS·Satoshi Inouye
Oct 11, 2011·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Vadim R VivianiFrederico G C Arnoldi
Feb 28, 2006·Luminescence : the Journal of Biological and Chemical Luminescence·Eileen Kim-ChoiIrina Rybina
Sep 16, 2011·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Luís Pinto da Silva, Joaquim C G Esteves da Silva
Jun 4, 2011·Journal of Computational Chemistry·Luís Pinto Da Silva, Joaquim C G Esteves Da Silva
Oct 30, 2009·Protein Science : a Publication of the Protein Society·Xueyan LiYoshihiro Ohmiya
Nov 26, 2009·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Chun-Gang MinJi-Kang Feng
May 19, 2010·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Chun-Gang MinChia-Chung Sun
Jun 2, 2007·Analytical Biochemistry·Hiroya FujiiAkihiko Tokida
Jun 30, 2004·Biophysical Chemistry·Anna SvantessonPål Nyrén
Sep 12, 2006·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Eileen Kim-ChoiIrina Rybina
Feb 9, 2006·Journal of Photochemistry and Photobiology. B, Biology·N S Rodionova, V N Petushkov
Sep 19, 2012·International Journal of Biological Macromolecules·Ali Riahi-MadvarFatemeh Rezaee
Jul 16, 2002·Annual Review of Biomedical Engineering·Christopher H Contag, Michael H Bachmann
Dec 27, 2011·Chemistry & Biology·Katryn R HarwoodStephen C Miller
Feb 28, 2009·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·V R VivianiF C Abdalla
Oct 31, 2002·Luminescence : the Journal of Biological and Chemical Luminescence·N N Ugarova, L Y Brovko
Jan 21, 2017·Journal of the American Chemical Society·Krysten A JonesJennifer A Prescher
Dec 12, 2017·ChemistryOpen·Michael C PirrungJiayu Liao

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