8IZ3

Single excitation and two emissions pH sensor protein(SITE-pHorin)_pH5.5

Summary for 8IZ3

• Entry DOI: 10.2210/pdb8iz3/pdb
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: monomer, fluorescent protein
• Biological source: Aequorea victoria (Water jellyfish, Mesonema victoria)
• Total number of polymer chains: 1
• Total formula weight: 27677.21

Authors

Kang, J.S.,Li, S.A. (deposition date: 2023-04-06, release date: 2024-10-09, Last modification date: 2025-10-22)

Primary citation

Li, S.A.,Meng, X.Y.,Zhang, S.,Zhang, Y.J.,Yang, R.Z.,Wang, D.D.,Yang, Y.,Liu, P.P.,Kang, J.S.
A unified intracellular pH landscape with SITE-pHorin: a quantum-entanglement-enhanced pH probe.
Sci China Life Sci, 2025

PubMed Abstract: An accurate map of intracellular organelle pH is crucial for comprehending cellular metabolism and organellar functions. However, a unified intracellular pH spectrum using a single probe is still lacking. Here, we developed a novel quantum entanglement-enhanced pH- sensitive probe called SITE-pHorin (single excitation and two emissions pH sensor protein), which features a wide pH-sensitive range and ratiometric quantitative measurement capabilities. We subsequently measured the pH of various organelles and their subcompartments, including mitochondrial subspaces, Golgi stacks, endoplasmic reticulum (ER), lysosomes, peroxisomes, and endosomes in COS-7 cells. For the long-standing debate on the pH of the mitochondrial compartments, we measured the pH of the mitochondrial cristae (mito-cristae) as 6.60±0.40, the pH of the mitochondrial intermembrane space (mito-IMS) as 6.95±0.30, and the pH of the two populations of the mitochondrial matrix (mito-matrix) at approximately 7.20±0.27 and 7.50±0.16, respectively. Notably, the pH of the lysosome exhibited a single, narrow Gaussian distribution centered at 4.79±0.17, which is consistent with an optimal lysosomal acidic pH between 4.5 and 5.0. Furthermore, quantum chemistry computations revealed that both the deprotonation of the residue Y182 and the discrete curvature of the deformed benzene ring in the chromophore are necessary for the quantum entanglement mechanism of SITE-pHorin. Intriguingly, our findings reveal an accurate pH gradient (0.6-0.9 pH units) between the mitochondrial cristae and the mitochondrial matrix, suggesting that prior knowledge about ΔpH (0.4-0.6) and the mitochondrial proton motive force (pmf) is underestimated.

PubMed: 40956361
DOI: 10.1007/s11427-025-2971-5

Experimental method

X-RAY DIFFRACTION (2.18 Å)