6AA6

X-ray structure of ReQy1 (reduced form)

Summary for 6AA6

• Entry DOI: 10.2210/pdb6aa6/pdb
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: yfp, fluorescent indicator protein, redox, fluorescent protein
• Biological source: Aequorea victoria (Jellyfish)
• Total number of polymer chains: 2
• Total formula weight: 56751.96

Authors

Kurisu, G.,Yasuda, A.,Tabushi, N.,Tanaka, H. (deposition date: 2018-07-17, release date: 2019-05-29, Last modification date: 2023-11-22)

Primary citation

Sugiura, K.,Tanaka, H.,Kurisu, G.,Wakabayashi, K.I.,Hisabori, T.
Multicolor redox sensor proteins can visualize redox changes in various compartments of the living cell.
Biochim Biophys Acta Gen Subj, 1863:1098-1107, 2019

PubMed Abstract: Change in the intracellular redox state is a consequence of various metabolic reactions, which simultaneously regulates various physiological phenomena in cells. Monitoring the redox state in living cells is thus very important for understanding cellular physiology. Various genetically encoded fluorescent redox sensors have therefore been developed. Recently, we developed oxidation-sensitive fluorescent proteins named Oba-Q (Sugiura, K., et al. (2015) Biochem. Biophys. Res. Commun. 457, 242-248), which exhibit dramatic quenching under oxidizing conditions. To extend the range of uses of redox sensor proteins, we refined these proteins based on the molecular architecture applied to Oba-Q, and successfully produced several redox sensor proteins based on CFP and YFP. Interestingly, some of these sensor proteins showed the reverse changes in emission compared with Oba-Q, implying remarkable fluorescence quenching under reducing conditions. We named this type of sensor protein Re-Q, reduction-sensed quenching protein. The cause of the redox-dependent fluorescence quenching could be clearly explained based on the crystal structure of Re-Q in the reduced and oxidized forms. In addition, by introducing suitable mutations into the sensors, we produced Oba-Q and Re-Q mutants exhibiting various midpoint redox potentials. This series of proteins can cover a wide range of redox potentials in the cell, so they should be applicable to various cells and even intracellular organelles. As an example, we successfully measured the redox responses in different cell compartments of cultured mammalian cells simultaneously against the anticancer reagents Kp372-1.

PubMed: 30953671
DOI: 10.1016/j.bbagen.2019.01.016

Experimental method

X-RAY DIFFRACTION (2.39 Å)