Application of new semisynthetic aequorins with long half-decay time of luminescence to G-protein-coupled receptor assay

作者：Inouye Satoshi^*; Iimori Rie; Sahara Yuiko; Hisada Sunao; Hosoya Takamitsu

来源：Analytical Biochemistry, 2010, 407(2): 247-252.

DOI：10.1016/j.ab.2010.08.025

• Aequorin
• Coelenterazine synthesis
• Cell-based GPCR assay
• High-throughput screening
• Fluorescent protein

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.

• 出版日期2010-12-15