Application of new semisynthetic aequorins with long half-decay time of luminescence to G-protein-coupled receptor assay.
Aequorin is a Ca(2+)-binding photoprotein and consists of an apoprotein (apoaequorin) and a 2-peroxide of coelenterazine. Eight new coelenterazine analogues modified at the C2-position were synthesized and incorporated into recombinant apoaequorin with O(2) to yield different semisynthetic aequorins. The luminescence properties and the sensitivity to Ca(2+) of these semisynthetic aequorins were characterized. Two semisynthetic aequorins, namely me- and cf3-aequorin, showed a slow decay of the luminescence pattern with less sensitivity to Ca(2+) and were useful for the cell-based G-protein-coupled receptor (GPCR) reporter assays.
Inouye
Satoshi
S
Yokohama Research Center, Chisso Co., 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. sinouye@chisso.co.jp
Iimori
Rie
R
Sahara
Yuiko
Y
Hisada
Sunao
S
Hosoya
Takamitsu
T
D000331:Aequorin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002118:Calcium / Q000737:chemistry; D006224:Cricetinae; D003412:Cricetulus; D006207:Half-Life; D007093:Imidazoles / Q000138:chemical synthesis / Q000737:chemistry; D007700:Kinetics; D049408:Luminescent Agents / Q000737:chemistry*; D010100:Oxygen / Q000737:chemistry; D011719:Pyrazines / Q000138:chemical synthesis / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D011976:Receptors, Muscarinic / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*
D001059:Apoproteins; D007093:Imidazoles; D049408:Luminescent Agents; D011719:Pyrazines; D043562:Receptors, G-Protein-Coupled; D011976:Receptors, Muscarinic; D011994:Recombinant Proteins; C060014:apoaequorin; C017144:coelenterazine; D000331:Aequorin; D010100:Oxygen; D002118:Calcium