γ-Oxo-1-pyrenebutyric acid used for fluorescent detection of serum albumins and trypsin

Abstract

Fluorescence spectroscopy, one of the most informative analytical techniques, has played and continues to play a key role in modern research due to its high sensitivity, rapid response rate, and relatively low cost. Herein we report its application to the detection of the proteins bovine serum albumin (BSA) and human serum albumin (HSA) as well as a protease (trypsin). The detection is based on a fluorescent molecule: c-oxo-1-pyrenebutyric acid (OPBA), which exhibits a quenched fluorescent change at 455 nm toward serum albumins (SAs). OPBA interacted with SAs with a 1 : 1 stoichiometry and a strong affinity due to p-stacking, hydrophobic interactions, and hydrogen bonding interactions. The microenvironment created by HSA and BSA played an important role in their respective interactions with OPBA. In addition, OPBA can be used for monitoring trypsin by the cleavage of HSA or BSA in the presence of copper ions, and the successful cleavage of SAs was demonstrated by the SDS polyacrylamide gel electrophoresis (PAGE) results. This indirect detection of trypsin might be developed as a strategy for sensors without substrate selection. All of these tests do not require sophisticated instrumentation and should be applicable to standard fluorescence assays.

Interaction of 1-pyrene sulfonic acid sodium salt with human serum albumin

“…Pyrene probes have strong absorption bands in the UV region, a high fluorescence quantum yield and long life time of the excited singlet state (can be examined with a nanosecond flash photolysis system). However, the literature concerning binding of pyrene and its derivatives to HSA or other serum albumins is not extensive [13,[19][20][21][22][23][24][25]. The albumin-induced changes of absorption and fluorescence spectra have been reported, but in most cases the authors do not specify unambiguously the probe/protein binding sites.…”

A selective fluorescence quenching method for the determination of trace hypochlorite in water samples with nile blue A

“…Fluorescence quenching includes static and dynamic quenching [25]. The quenching mechanism can be distinguished by their differing dependence on temperature.…”

Study on the Interaction between Rhodamine Dyes and Allura Red Based on Fluorescence Spectra and Its Analytical Application in Soft Drinks

“…In addition, the quenching mechanism, including dynamic quenching and static quenching. 31 Firstly, the characteristics of the UV-visible absorption spectra of AR/R6G/AR-R6G were investigated to explore the quenching mechanism. As can be observed from Fig.…”