title: Detection of zinc finger protein (EGR1) based on electrogenerated chemiluminescence from singlet oxygen produced in a nanoclay-supported porphyrin environment.
abstract: Early growth response protein 1 (EGR1), as a characteristic example of zinc finger proteins, acts as a transcription factor in eukaryotic cells, mediating protein-protein interactions. Here, a novel electrochemiluminescence (ECL)-based protocol for EGR1 assay was developed with a new eco-friendly emitter: singlet oxygen produced in the vicinity of nanoclay-supported zinc proto-porphyrin IX (ZnPPIX). Its electrochemical reduction stimulates an intense monochromic CL irradiation at 644 nm from the dissolved oxygen as endogenous coreactant in the aqueous solution. This ECL derivation was rationalized via hyphenated spectroscopy and theoretical calculation. To promote hydrophilicity and solid-state immobilization of porphyrins, the lamellar artificial laponite was employed as a nanocarrier owning to its large specific area without the blackbody effect. The facile exfoliation of laponite produced quality monolayered nanosheets and facilitated the adsorption and flattening of PPIX upon the surface, resulting in a highly efficient ECL emission. Based on the release of Zn(2+) in zinc finger domains of EGR1 upon contact with the ECL-inactive PPIX, which was monitored by circular dichroism and UV-absorption, a sensitive Zn(2+)-selective electrode for the "signal-on" detection of EGR1 was prepared with a detection limit down to 0.48 pg mL(-1) and a linearity over 6 orders of magnitude. The proposed porphyrin-based ECL system thus infused fresh blood into the traditional ECL family, showing great promise in bioassays of structural Zn(II) proteins and zinc finger-binding nucleotides.
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Shengyuan
authors: S
authors: Deng
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Tingting
authors: T
authors: Zhang
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Xubo
authors: X
authors: Ji
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.|Intelligent Microsystem Technology and Engineering Center, School of Mechanical Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Ying
authors: Y
authors: Wan
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Peng
authors: P
authors: Xin
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Dan
authors: D
authors: Shan
authors: Sino-French Laboratory of Biomaterials and Bioanalytical Chemistry, School of Environmental and Biological Engineering, Nanjing University of Science and Technology , Nanjing 210094, P. R. China.
authors: Xueji
authors: X
authors: Zhang
mesh_terms: D000538:Aluminum Silicates / Q000737:chemistry*; D000077215:Clay; D051766:Early Growth Response Protein 1 / Q000032:analysis* / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D007700:Kinetics; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008665:Metalloporphyrins / Q000737:chemistry*; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry; D017640:Silicates / Q000737:chemistry; D026082:Singlet Oxygen / Q000737:chemistry*; D015032:Zinc / Q000737:chemistry; D016335:Zinc Fingers
chemical_list: D000538:Aluminum Silicates; D051766:Early Growth Response Protein 1; D008665:Metalloporphyrins; D017640:Silicates; D026082:Singlet Oxygen; C524813:laponite; D015032:Zinc; D000077215:Clay