9U9D

Bipartite Genetically Encoded Biosensor sG-GECO1

Summary for 9U9D

• Entry DOI: 10.2210/pdb9u9d/pdb
• Descriptor: Myosin light chain kinase, smooth muscle, deglutamylated form,Green fluorescent protein,Calmodulin-1, Green fluorescent protein (3 entities in total)
• Functional Keywords: fluorescent protein, gfp, bipartite scpfp, sg-geco1
• Biological source: synthetic construct; More
• Total number of polymer chains: 2
• Total formula weight: 52850.17

Authors

Wen, Y.,Campbell, R.E. (deposition date: 2025-03-27, release date: 2025-09-17, Last modification date: 2025-10-01)

Primary citation

Yamaguchi, I.,Barazzuol, L.,Dematteis, G.,Zhu, W.,Wen, Y.,Drobizhev, M.,Lim, D.,Campbell, R.E.,Cali, T.,Nasu, Y.
Bipartite Genetically Encoded Biosensors to Sense Calcium Ion Dynamics at Membrane-Membrane Contact Sites.
Anal.Chem., 97:19848-19861, 2025

PubMed Abstract: Self-complementing bipartite fluorescent proteins (FPs) are useful tools for the detection of protein-protein proximity and for localizing fluorophores to membrane-membrane contact sites. Here, we report versions of circularly permuted green FP (GFP), red FP (RFP), and mNeonGreen (NG), which are split into a large fragment composed of nine β-strands and a small fragment composed of two β-strands. In each case, the large and small fragments can associate in live cells to form the complete 11-stranded FP β-barrel. We further converted each of these three self-complementing FPs into bipartite calcium ion (Ca) biosensors. We demonstrate that appropriately targeted versions of these split FPs, and split FP-based biosensors, can be functionally assembled at membrane-membrane contact sites. We employ the bipartite NG-based Ca biosensor for visualization of pharmacologically induced Ca release at mitochondria-endoplasmic reticulum contact sites (MERCs).

PubMed: 40888292
DOI: 10.1021/acs.analchem.5c03831

Experimental method

X-RAY DIFFRACTION (1.8 Å)