3SG4

Crystal Structure of GCaMP3-D380Y, LP(linker 2)

3SG4 の概要

• エントリーDOI: 10.2210/pdb3sg4/pdb
• 関連するPDBエントリー: 3SG2 3SG3 3SG5 3SG6 3SG7
• 分子名称: Myosin light chain kinase, Green fluorescent protein, Calmodulin-1 chimera, CALCIUM ION (3 entities in total)
• 機能のキーワード: calcium sensor, fluorescent protein
• 由来する生物種: Gallus gallus (Chicken); 詳細
• 細胞内の位置: Cytoplasm, cytoskeleton, spindle: P0DP29
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 50704.76

構造登録者

Schreiter, E.R.,Akerboom, J.,Looger, L.L. (登録日: 2011-06-14, 公開日: 2012-06-20, 最終更新日: 2024-10-16)

主引用文献

Akerboom, J.,Chen, T.W.,Wardill, T.J.,Tian, L.,Marvin, J.S.,Mutlu, S.,Calderon, N.C.,Esposti, F.,Borghuis, B.G.,Sun, X.R.,Gordus, A.,Orger, M.B.,Portugues, R.,Engert, F.,Macklin, J.J.,Filosa, A.,Aggarwal, A.,Kerr, R.A.,Takagi, R.,Kracun, S.,Shigetomi, E.,Khakh, B.S.,Baier, H.,Lagnado, L.,Wang, S.S.,Bargmann, C.I.,Kimmel, B.E.,Jayaraman, V.,Svoboda, K.,Kim, D.S.,Schreiter, E.R.,Looger, L.L.
Optimization of a GCaMP calcium indicator for neural activity imaging.
J.Neurosci., 32:13819-13840, 2012

PubMed Abstract: Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Recent efforts in protein engineering have significantly increased the performance of GECIs. The state-of-the art single-wavelength GECI, GCaMP3, has been deployed in a number of model organisms and can reliably detect three or more action potentials in short bursts in several systems in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro assays, we have increased the dynamic range of GCaMP3 by severalfold, creating a family of "GCaMP5" sensors. We tested GCaMP5s in several systems: cultured neurons and astrocytes, mouse retina, and in vivo in Caenorhabditis chemosensory neurons, Drosophila larval neuromuscular junction and adult antennal lobe, zebrafish retina and tectum, and mouse visual cortex. Signal-to-noise ratio was improved by at least 2- to 3-fold. In the visual cortex, two GCaMP5 variants detected twice as many visual stimulus-responsive cells as GCaMP3. By combining in vivo imaging with electrophysiology we show that GCaMP5 fluorescence provides a more reliable measure of neuronal activity than its predecessor GCaMP3. GCaMP5 allows more sensitive detection of neural activity in vivo and may find widespread applications for cellular imaging in general.

PubMed: 23035093
DOI: 10.1523/JNEUROSCI.2601-12.2012

実験手法

X-RAY DIFFRACTION (2.4 Å)