Fluorescent Probes and Quenchers in Studies of Protein Folding and Protein‐Lipid Interactions

Abstract

Fluorescence spectroscopy provides numerous methodological tools for structural and functional studies of biological macromolecules and their complexes. All fluorescence‐based approaches require either existence of an intrinsic probe or an introduction of an extrinsic one. Moreover, studies of complex systems often require an additional introduction of a specific quencher molecule acting in combination with a fluorophore to provide structural or thermodynamic information. Here, we review the fundamentals and summarize the latest progress in applications of different classes of fluorescent probes and their specific quenchers, aimed at studies of protein folding and protein‐membrane interactions. Specifically, we discuss various environment‐sensitive dyes, FRET probes, probes for short‐distance measurements, and several probe‐quencher pairs for studies of membrane penetration of proteins and peptides. The goals of this review are: (a) to familiarize the readership with the general concept that complex biological systems often require both a probe and a quencher to decipher mechanistic details of functioning and (b) to provide example of the immediate applications of the described methods.

Chemistry in Ukraine

“…Fluorescent probes and their specific quencher's applications to study protein folding and protein-membrane interactions are summarized in the progress review of Kyrychenko and Ladokhin. [17] The additional goal of the review is to familiarize the broad readership with the general concepts of deciphering mechanistic details of protein functioning and provide examples of their applications.…”

Unnatural Amino Acids for Biological Spectroscopy and Microscopy

“…Therefore, these intrinsic fluorescent amino acids have found a wide range of utilities in biological sciences. However, because this topic has been extensively reviewed in the past, − herein we only highlight a few recent examples where Trp is employed to study various biophysical questions.…”