A FRET-based Calcium Biosensor based on Calmodulin (CaM) and its newly identified binding peptide fr

来源 :中国化学会第三届全国生物物理化学会议暨国际华人生物物理化学发展论坛 | 被引量 : 0次 | 上传用户:j621212
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Genetically encoded calcium biosensors are valuable tools in cell biology and neuroscience.A FRET-based calcium biosensor Cameleon has received a great deal of optimization since it was released in 1997.Despite all the extensive improvements,the sensing and functional domains of Cameleon were solely based on engineering of the fusion protein CaM and its binding peptide from myosin light-chain kinase (M13).Whether there exist other feasible CaM-binding peptides that can replace M13 in sensor construction has not been explored.Here,we report a FRET-based calcium biosensor comprising CaM and its newly identified binding peptide from c-FLIPL.We tested the performance of this new CaM-binding peptide from c-FLIPL in construction of functional FRET-based biosensors.We also performed preliminary optimization and application in imaging of Ca2+ dynamics in living cells.
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