In Vitro Light‐Up Visualization of a Subunit‐Specific Enzyme by an AIE Probe via Restriction of Single Molecular Motion

Abstract

Enzymes contain several subunits to maintain different biological functions. However, it remains a great challenge for specific discrimination of one subunit over another. Toward this end, the fluorescent probe TPEMA is now presented for highly specific detection of the B subunit of cytosolic creatine (CK) kinase isoenzyme (CK‐B). Owing to its aggregation‐induced emission property, TPEMA shows highly boosted emission toward CK‐B with a fast response time and very low interference from other analytes, including the M subunit of CK (CK‐M). With the aid of a Job plot assay, ITC assay and molecular dynamics simulation, it was directly confirmed that the remarkably enhanced fluorescence of TPEMA in the presence of CK‐B results from the restriction of single molecular motion in the cavity. Selective wash‐free fluorescence imaging of CK‐B in macrophages under different treatments was successfully demonstrated.

A Bio‐Conjugated Fullerene as a Subcellular‐Targeted and Multifaceted Phototheranostic Agent

“…Due to the insolubility of C 70 in water and water‐miscible solvents, standard methodologies for determining binding energy, such as ITC or fluorescence titration analysis, [ 67 ] cannot be performed for C 70 and lysozyme. Nevertheless, we determined the binding affinity between the two interacting systems using a computational protocol, which was recently validated to calculate the binding energy between proteins and fullerenes (Section S7, Supporting Information).…”

Fluorescent probes for visualizing ROS-associated proteins in disease

“…34 ). 136 The inhibitor ethylmalonic acid was used as the targeting moiety to deliver the probe into the cavity of CK-B, and TPE was used as the fluorophore. When probe 46 was inserted into the cavity of CK-B, the molecular motion of TPE was restricted, and TPE emitted strong fluorescence.…”

Recent Developments in Fluorescent Materials for Heavy Metal Ions Analysis From the Perspective of Forensic Chemistry

“…There are a wide array of instrumental methods allowing identification and determination of various metal elements in different evidences, such as atomic absorption spectroscopy, neutron activation analysis, X-ray fluorescence spectroscopy, ion chromatography, mass spectroscopy, and other spectroscopy coupled with mass spectroscopy (Ulrich et al, 2004;Verma, 2018;Sliwińska et al, 2019). Compared with these traditional analytical methods, fluorescence spectroscopy is more suitable for trace metal ions analysis in complex matrices because of its short response time, simplicity, high selectivity, and sensitivity (Fu et al, 2019;Zhang et al, 2019;Niu et al, 2020;Zang et al, 2020).…”