Deciphering the molecular mechanism responsible for GCaMP6m's Ca2+-dependent change in fluorescence

被引：91

作者：

Barnett, Lauren M. ^{[1
]}

Hughes, Thomas E. ^{[1
]}

Drobizhev, Mikhail ^{[1
]}

机构：

[1] Montana State Univ, Dept Cell Biol & Neurosci, Bozeman, MT 59717 USA

来源：

PLOS ONE | 2017年 / 12卷 / 02期

关键词：

2-PHOTON ABSORPTION PROPERTIES; IMAGING NEURAL ACTIVITY; CALCIUM INDICATOR; STRUCTURAL BASIS; PROTEIN; STATE; PROBE; PH; CHROMOPHORE; ACTIVATION;

D O I：

10.1371/journal.pone.0170934

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

The goal of this work is to determine how GCaMP6m's fluorescence is altered in response to Ca2+-binding. Our detailed spectroscopic study reveals the simplest explanation for how GCaMP6m changes fluorescence in response to Ca2+ is with a four-state model, in which a Ca2+-dependent change of the chromophore protonation state, due to a shift in pK(a), is the predominant factor. The pKa shift is quantitatively explained by a change in electrostatic potential around the chromophore due to the conformational changes that occur in the protein when calmodulin binds Ca2+ and interacts with the M13 peptide. The absolute pKa values for the Ca2+-free and Ca2+-saturated states of GCaMP6m are critical to its high signal-to-noise ratio. This mechanism has important implications for further improvements to GCaMP6m and potentially for other similarly designed biosensors.

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