Inhaled Anesthetic Binding Sites in Human Serum Albumin

Previous evidence suggests multiple anesthetic binding sites on human serum albumin, but to date, we have only identified Trp-214 in an interdomain cleft as contributing to a binding site. We used a combination of site-directed mutagenesis, photoaffinity labeling, amide hydrogen exchange, and trypto...

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Bibliographic Details
Published in:The Journal of biological chemistry 2000-09, Vol.275 (39), p.30439-30444
Main Authors: Eckenhoff, Roderic G., Petersen, Charles E., Ha, Chung-Eun, Bhagavan, Nadhipuram V.
Format: Article
Language:English
Subjects:
Anesthetics, Inhalation - metabolism
Binding Sites
Cyclobutanes - metabolism
Halothane - metabolism
Humans
Isoflurane - metabolism
Models, Molecular
Mutagenesis, Site-Directed
Peptide Fragments - metabolism
Protein Conformation
Serum Albumin - chemistry
Serum Albumin - genetics
Serum Albumin - metabolism
Spectrometry, Fluorescence
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Summary:Previous evidence suggests multiple anesthetic binding sites on human serum albumin, but to date, we have only identified Trp-214 in an interdomain cleft as contributing to a binding site. We used a combination of site-directed mutagenesis, photoaffinity labeling, amide hydrogen exchange, and tryptophan fluorescence spectroscopy to evaluate the importance to binding of a large domain III cavity and compare it to binding character of the 214 interdomain cleft. The data show anesthetic binding in this domain III cavity of similar character to the interdomain cleft, but selectivity for different classes of anesthetics exists. Occupancy of these sites stabilizes the native conformation of human serum albumin. The features necessary for binding in the cleft appear to be fairly degenerate, but in addition to hydrophobicity, there is evidence for the importance of polarity. Finally, myristate isosterically competes with anesthetic binding in the domain III cavity and allosterically enhances anesthetic binding in the interdomain cleft.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M005052200