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Cited by 28Protein Interaction Studies and Fluorescence AnalysisProtein Structure and DynamicsDrug Transport and Resistance Mechanisms

Allosterically Coupled Multisite Binding of Testosterone to Human Serum Albumin

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Jayaraj, Abhilash, Schwanz, Heidi A, Spencer, Daniel J, Bhasin, Shalender, Hamilton, James A, Jayaram, B, Goldman, Anna L, Krishna, Meenakshi +12 more·2021·Endocrinology
DOI 10.1210/endocr/bqaa199PMC PMC7774055
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Abstract

Human serum albumin (HSA) acts as a carrier for testosterone, other sex hormones, fatty acids, and drugs. However, the dynamics of testosterone’s binding to HSA and the structure of its binding sites remain incompletely understood. Here, we characterize the dynamics of testosterone’s binding to HSA and the stoichiometry and structural location of the binding sites using 2-dimensional nuclear magnetic resonance (2D NMR), fluorescence spectroscopy, 4,4′-dianilino-1,1′-binaphthyl-5,5′-disulfonic acid dipotassium salt partitioning, and equilibrium dialysis, complemented by molecular modeling. 2D NMR studies showed that testosterone competitively displaced 18-[13C]-oleic acid from at least 3 known fatty acid binding sites on HSA that also bind many drugs. Binding isotherms of testosterone’s binding to HSA generated using fluorescence spectroscopy and equilibrium dialysis were nonlinear and the apparent dissociation constant varied with different concentrations of testosterone and HSA. The binding isotherms neither conformed to a linear binding model with 1:1 stoichiometry nor to 2 independent binding sites; the binding isotherms were most consistent with 2 or more allosterically coupled binding sites. Molecular dynamics studies revealed that testosterone’s binding to fatty acid binding site 3 on HSA was associated with conformational changes at site 6, indicating that residues in in these 2 distinct binding sites are allosterically coupled. There are multiple, allosterically coupled binding sites for testosterone on HSA. Testosterone shares these binding sites on HSA with free fatty acids, which could displace testosterone from HSA under various physiological states or disease conditions, affecting its bioavailability.

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  • Binding site competition and allosteric coupling can, in conditions such as postprandial and heightened fatty acids and drug usage, result in displacement of testosterone from its binding site, potentially affecting testosterone bioavailability and/or metabolism.

    “Binding site competition and allosteric coupling can in certain conditions, especially postprandial and in circumstances of heightened levels of fatty acids and drug usage, result in displacement of T from its binding site thereby potentially affecting T bioavailability and/or metabolism [25, 26].”

    From Role of sex hormone-binding globulin in the free hormone hypothesis and the relevance of free testosterone in androgen physiology (2022) · cites this paper as [25]

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Authors
Jayaraj, A., Schwanz, H. A., Spencer, D. J., Bhasin, S., Hamilton, J. A., Jayaram, B., Goldman, A. L., Krishna, M., Krishnan, M., Shah, A., Jin, Z., Krenzel, E., Nair, S. N., Ramesh, S., Guo, W., Wagner, G., Arthanari, H., Peng, L., Lawney, B., & Jasuja, R.
Affiliations
  • Indian Institute of Technology Delhi (IN)
  • Boston University (US)
  • Brigham and Women's Hospital (US)
  • University of Iowa (US)
  • Harvard University (US)
Corresponding author
Jasuja, Ravi
ORCID
Venue
Endocrinology
Publisher
The Endocrine Society
Published
2020-11-30
Volume
162
Issue
2
ISSN
  • 0013-7227 (ISSN-L)
  • 1945-7170
DOI
10.1210/endocr/bqaa199
PMC
PMC7774055
OpenAlex
W3096778261
Cited by
28
References
4
Citations by year
2026: 5 · 2025: 2 · 2024: 7 · 2023: 3 · 2022: 7 · 2021: 4
FWCI
1.13
Type
journal article
Language
en
Open access
Licence
academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
Locations
  • Endocrinology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
National Institutes of Health (R43AG045011, R44AG045011)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
testosterone, human serum albumin, allosteric communication, binding stoichiometry, binding isotherms, fatty acid binding sites, site 3, conformational change, drug binding, steroid binding, bioavailability
MeSH terms
Serum Albumin, Human, Carbon Isotopes, Magnetic Resonance Spectroscopy, Spectrometry, Fluorescence, Testosterone, Molecular Dynamics Simulation

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