Faculty Publications
Document Type
Article
Publication Version
Published Version
Journal/Book/Conference Title
Biochemistry
Volume
63
Issue
1
First Page
19
Last Page
26
Abstract
The cholesterol affinities of many integral plasma membrane proteins have been estimated by molecular computation. However, these values lack experimental confirmation. We therefore developed a simple mathematical model to extract sterol affinity constants and stoichiometries from published isotherms for the dependence of the activity of such proteins on the membrane cholesterol concentration. The binding curves for these proteins are sigmoidal, with strongly lagged thresholds attributable to competition for the cholesterol by bilayer phospholipids. The model provided isotherms that matched the experimental data using published values for the sterol association constants and stoichiometries of the phospholipids. Three oligomeric transporters were found to bind cholesterol without cooperativity, with dimensionless association constants of 35 for Kir3.4* and 100 for both Kir2 and a GAT transporter. (The corresponding ΔG° values were −8.8, −11.4, and −11.4 kJ/mol, respectively). These association constants are significantly lower than those for the phospholipids, which range from ∼100 to 6000. The BK channel, the nicotinic acetylcholine receptor, and the M192I mutant of Kir3.4* appear to bind multiple cholesterol molecules cooperatively (n = 2 or 4), with subunit affinities of 563, 950, and 700, respectively. The model predicts that the three less avid transporters are approximately half-saturated in their native plasma membranes; hence, they are sensitive to variations in cholesterol in vivo. The more avid proteins would be nearly saturated in vivo. The method can be applied to any integral protein or other ligands in any bilayer for which there are reasonable estimates of the sterol affinities and stoichiometries of the phospholipids.
Department
Department of Physics
Original Publication Date
1-2-2024
Object Description
1 PDF File
DOI of published version
10.1021/acs.biochem.3c00567
Repository
UNI ScholarWorks, Rod Library, University of Northern Iowa
Copyright
©2023 The Authors. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Language
en
File Format
application/pdf
Recommended Citation
Steck, Theodore L.; Ali Tabei, S. M.; and Lange, Yvonne, "Estimating the Cholesterol Affinity of Integral Membrane Proteins from Experimental Data" (2024). Faculty Publications. 5638.
https://scholarworks.uni.edu/facpub/5638
Comments
First published in Biochemistry, v63 i1 published by American Chemistry Society. DOI: https://doi.org/10.1021/acs.biochem.3c00567