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Resolving Binding Events on the Multifunctional Human Serum Albumin
Author(s) -
Wenskowsky Lea,
Wagner Michael,
Reusch Johannes,
Schreuder Herman,
Matter Hans,
Opatz Till,
Petry Stefan Matthias
Publication year - 2020
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.202000069
Subject(s) - plasma protein binding , binding site , chemistry , human serum albumin , biophysics , biomolecule , receptor–ligand kinetics , computational biology , drug discovery , serum albumin , biochemistry , biology , receptor
Physiological processes rely on initial recognition events between cellular components and other molecules or modalities. Biomolecules can have multiple sites or mode of interaction with other molecular entities, so that a resolution of the individual binding events in terms of spatial localization as well as association and dissociation kinetics is required for a meaningful description. Here we describe a trichromatic fluorescent binding‐ and displacement assay for simultaneous monitoring of three individual binding sites in the important transporter and binding protein human serum albumin. Independent investigations of binding events by X‐ray crystallography and time‐resolved dynamics measurements (switchSENSE technology) confirm the validity of the assay, the localization of binding sites and furthermore reveal conformational changes associated with ligand binding. The described assay system allows for the detailed characterization of albumin‐binding drugs and is therefore well‐suited for prediction of drug‐drug and drug‐food interactions. Moreover, conformational changes, usually associated with binding events, can also be analyzed.