Kim, Tae-Jin Kim, Kyung-A Jung, Sang Hoon 2024-01-20T01:02:07Z 2024-01-20T01:02:07Z 2021-09-04 2017-08 0925-4005 https://pubs.kist.re.kr/handle/201004/122498 Endoplasmic reticulum (ER) is an organelle critically involved in Ca2+ signaling by regulating the intracellular Ca2+ gradient. In the present study, we have developed an ER Ca2+ sensor based on fluorescence resonance energy transfer (FRET), capable of monitoring dynamic Ca2+ signals within the ER. This genetically-encoded ER sensor contains a mutated version of calmodulin (mCaM) and its binding peptide derived from smooth muscle myosin light chain kinase (m-smMLCK) as the Ca2+-responsive elements, thereby enabling an effective ER Ca2+ sensing without perturbation by endogenous CaM. TheCa(2+) sensitivity, including detection limit, of the sensor was evaluated through in vitro characterization. In cell-based assay, our biosensor displayed high accuracy to ER Ca2+ sensing in response to pharmacological stimuli, indicating that this sensor is reliable for visualization of ER Ca2+ signals at high spatiotemporal resolutions in live cells. (C) 2017 Elsevier B.V. All rights reserved. English ELSEVIER SCIENCE SA GENE-EXPRESSION CA2+ INDICATORS CALMODULIN PROTEINS DYNAMICS RELEASE PATHWAY INDO-1 Development of an endoplasmic reticulum calcium sensor based on fluorescence resonance energy transfer Article 10.1016/j.snb.2017.03.083 1 SENSORS AND ACTUATORS B-CHEMICAL, v.247, pp.520 - 525 SENSORS AND ACTUATORS B-CHEMICAL 247 520 525 scie scopus 000402465000063 2-s2.0-85015877400 Chemistry, Analytical Electrochemistry Instruments & Instrumentation Chemistry Electrochemistry Instruments & Instrumentation Article GENE-EXPRESSION CA2+ INDICATORS CALMODULIN PROTEINS DYNAMICS RELEASE PATHWAY INDO-1 Fluorescence resonance energy transfer Calcium sensor Endoplasmic reticulum Live cell imaging