Leonardo Almeida Barison , Maurício Ikeda Yoguim , Aguinaldo Robinson de Souza , Nelson Henrique Morgon , Valdecir Farias Ximenes
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引用次数: 0
Abstract
AMPPD is an abbreviation for 3-(2′-spiroadamantyl)-4-methoxy-4-(3-phosphoryloxy)-phenyl-1,2-dioxetane, a thermally stable 1,2-dioxetane derivative widely used as a chemiluminescent probe in clinical laboratories for enzyme immunoassays. AMPPD is a substrate for alkaline phosphatase (ALP), the enzyme that catalyzes the remotion of the phosphoryl group of AMPPD, triggering light emission. This study investigated the impact of albumins on the AMPPD:ALP system and found that human serum albumin (HSA) increases the light emission efficiency of AMPPD:ALP by around 100 times. The efficiency of light enhancement varied depending on the type of albumin, with bovine (BSA), porcine (PSA), and rabbit (RSA) serum albumins being less effective than HSA. We conducted experiments to demonstrate that once hydrolyzed by ALP, the dephosphorylated AMPPD diffuses to site I of HSA. By studying the photophysical properties of the phenol ester generated after AMPPD cleavage, we found that its complexation with HSA caused a significant increase in its fluorescent efficiency, which explains the amplification of the chemiluminescence. Resonance energy transfer from the excited phenol ester to coumarin-153 was detected, confirming the presence of both molecules in the protein cavity. Hence, introducing fluorescent acceptors into the reaction system can adjust the emission wavelength. Besides the first direct evidence of albumin as an enhancer for chemiluminescence, the results highlight the potential interference of albumins on AMPPD-mediated assays and new biotechnological applications.
期刊介绍：
JPPA publishes the results of fundamental studies on all aspects of chemical phenomena induced by interactions between light and molecules/matter of all kinds.
All systems capable of being described at the molecular or integrated multimolecular level are appropriate for the journal. This includes all molecular chemical species as well as biomolecular, supramolecular, polymer and other macromolecular systems, as well as solid state photochemistry. In addition, the journal publishes studies of semiconductor and other photoactive organic and inorganic materials, photocatalysis (organic, inorganic, supramolecular and superconductor).
The scope includes condensed and gas phase photochemistry, as well as synchrotron radiation chemistry. A broad range of processes and techniques in photochemistry are covered such as light induced energy, electron and proton transfer; nonlinear photochemical behavior; mechanistic investigation of photochemical reactions and identification of the products of photochemical reactions; quantum yield determinations and measurements of rate constants for primary and secondary photochemical processes; steady-state and time-resolved emission, ultrafast spectroscopic methods, single molecule spectroscopy, time resolved X-ray diffraction, luminescence microscopy, and scattering spectroscopy applied to photochemistry. Papers in emerging and applied areas such as luminescent sensors, electroluminescence, solar energy conversion, atmospheric photochemistry, environmental remediation, and related photocatalytic chemistry are also welcome.
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