title: Titration of recombinant aequorin with calcium chloride.
abstract: The photoprotein aequorin emits light in the presence of a trace of Ca2+. The primary structure of the protein indicates the presence of three Ca2+-binding sites, whereas the luminometric titration of heterogeneous natural aequorin with Ca2+ has shown that the light emission takes place by the binding of two Ca2+ ions. In the case of recombinant aequorin, which is more suitable for quantitative studies, the titration with Ca2+ monitored by a Ca2+-sensitive electrode revealed that the photoprotein can bind more than two, most likely three, Ca2+ ions, and the luminometric titration conclusively showed that the luminescence is triggered by the first two Ca2+ ions bound. The affinity of recombinant aequorin for the first two Ca2+ ions, which are essential for light emission, was about 20 times stronger than that for the third Ca2+ ion, which is unrelated to light emission.
authors.affiliation: Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
authors.forename: O
authors.initials: O
authors.lastname: Shimomura
authors.forename: S
authors.initials: S
authors.lastname: Inouye
mesh_terms: D000331:Aequorin / Q000737:chemistry*; D001059:Apoproteins / Q000737:chemistry; D001665:Binding Sites; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D001059:Apoproteins; D011994:Recombinant Proteins; C060014:apoaequorin; D000331:Aequorin