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A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening
Published on: October 16, 2015
Inhaled anesthetic binding sites in human serum albumin
R G Eckenhoff1, C E Petersen, C E Ha
1Department of Anesthesia, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104-4283, USA. reckenho@mail.med.upenn.edu
Researchers discovered a new anesthetic binding site in human serum albumin (HSA) domain III, distinct from the previously known Trp-214 site. This finding reveals differential anesthetic selectivity and stabilization of HSA conformation.
Area of Science:
- Biochemistry
- Pharmacology
- Structural Biology
Background:
- Multiple anesthetic binding sites are suggested on human serum albumin (HSA).
- Trp-214 in an interdomain cleft was the only previously identified binding site.
- The role of a large domain III cavity in anesthetic binding remained unclear.
Purpose of the Study:
- To evaluate the importance of the domain III cavity for anesthetic binding.
- To compare the binding characteristics of the domain III cavity with the Trp-214 interdomain cleft.
- To investigate the influence of anesthetic binding on HSA conformation.
Main Methods:
- Site-directed mutagenesis
- Photoaffinity labeling
- Amide hydrogen exchange
- Tryptophan fluorescence spectroscopy
Main Results:
- Anesthetic binding occurs in the domain III cavity with characteristics similar to the interdomain cleft.
- Differential selectivity for anesthetic classes was observed between the two binding sites.
- Myristate competes with anesthetic binding in the domain III cavity and enhances it in the interdomain cleft.
Conclusions:
- HSA possesses multiple distinct anesthetic binding sites, including a significant cavity in domain III.
- Anesthetic binding to these sites stabilizes the native conformation of HSA.
- Binding site features involve hydrophobicity and polarity, with distinct interactions for different anesthetic classes.
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