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Influence of Domain Stability on the Properties of Human Apolipoprotein E3 and E4 and Mouse Apolipoprotein E

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Nguyen, David, Dhanasekaran, Padmaja, Nickel, Margaret, Mizuguchi, Chiharu, Watanabe, Mayu, Saito, Hiroyuki, Phillips, Michael C., Lund-Katz, Sissel·2014·Biochemistry
DOI 10.1021/bi500340zPMC PMC4071092
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Abstract

The human apolipoprotein (apo) E4 isoform, which differs from wild-type apoE3 by the single amino acid substitution C112R, is associated with elevated risk of cardiovascular and Alzheimer’s diseases, but the molecular basis for this variation between isoforms is not understood. Human apoE is a two-domain protein comprising an N-terminal helix bundle and a separately folded C-terminal region. Here, we examine the concept that the ability of the protein to bind to lipid surfaces is influenced by the stability (or readiness to unfold) of these domains. The lipid-free structures and abilities to bind to lipid and lipoprotein particles of a series of human and mouse apoE variants with varying domain stabilities and domain–domain interactions are compared. As assessed by urea denaturation, the two domains are more unstable in apoE4 than in apoE3. To distinguish the contributions of the destabilization of each domain to the greater lipid-binding ability of apoE4, the properties of the apoE4 R61T and E255A variants, which have the same helix bundle stabilities but altered C-terminal domain stabilities, are compared. In these cases, the effects on lipid-binding properties are relatively minor, indicating that the destabilization of the helix bundle domain is primarily responsible for the enhanced lipid-binding ability of apoE4. Unlike human apoE, mouse apoE behaves essentially as a single domain, and its lipid-binding characteristics are more similar to those of apoE4. Together, the results show that the overall stability of the entire apoE molecule exerts a major influence on its lipid- and lipoprotein-binding properties.

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  • The receptor-binding sites (AA136-150) for LDLR, LRP1, and HSPGs are located in a region conserved between mouse and human APOE.

    “However, the receptor-binding sites (AA136-150) for lipoprotein receptors such as the low-density lipoprotein receptor (LDLR), lipoprotein-related receptor 1 (LRP1), and heparan sulfate proteoglycans (HSPGs) are located in a region conserved between species (Fig. 1a) [11–14].”

    From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [11]

    Results > Structural Characterization (page unavailable) reports that human and mouse apoE sequences are 70% identical, supporting broad conservation between the species. It does not identify residues 136–150 as conserved or locate LDLR, LRP1, and HSPG binding sites there; searches for those residues and receptor-binding terms found no supporting passage. (LLM, conf=0.78)

    Experimental — accuracy not yet measured

  • Mouse APOE shares 70% homology with human APOE and is encoded by one major allele that resembles human APOE4 by having arginine at positions 112 and 158.

    “By contrast, mouse APOE (mAPOE) shares 70% homology with human APOE and is encoded by one major allele, which resembles the human APOE4 isoform by having arginine at both positions 112 and 158 [11].”

    From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [11]

    The Results section, under “Primary Structures of Human and Mouse ApoE Variants” (page unavailable), says the human and mouse sequences are 70% identical. The paper also says human apoE4 has arginine at positions 112 and 158, but our copy does not state that mouse apoE has those residues or is encoded by one major allele. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

  • mAPOE and hAPOE have a conserved LDLR and HSPG binding motif between AA136 to 150 (human numbering), which includes R136S Christchurch variant.

    “However, mAPOE and hAPOE have a conserved LDLR and HSPG binding motif between AA136 to 150 (human numbering) [11, 12, 39], which includes the R136S Christchurch variant.”

    From APOE Christchurch enhances a disease-associated microglial response to plaque but suppresses response to tau pathology (2025) · cites this paper as [11]

    Results > Structural Characterization reports 70% overall sequence identity between human and mouse apoE, but does not identify a conserved binding motif at residues 136–150. Searches for LDLR, HSPG, and the R136S Christchurch variant found no passage addressing that specific claim. No page number is available in our copy. (LLM, conf=0.72)

    Experimental — accuracy not yet measured

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Cites

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Authors
Nguyen, D., Dhanasekaran, P., Nickel, M., Mizuguchi, C., Watanabe, M., Saito, H., Phillips, M. C., & Lund-Katz, S.
Affiliations
  • Children's Hospital of Philadelphia (US)
  • Tokushima University (JP)
  • Lipid Research Group, Division of Gastroenterology, Hepatology and Nutrition, The Children’s Hospital of Philadelphia, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania 19104-4318, United States
ORCID
Venue
Biochemistry
Publisher
American Chemical Society (ACS)
Published
2014-06-11
Volume
53
Issue
24
Pages
4025-4033
ISSN
  • 0006-2960 (ISSN-L)
  • 1520-4995
DOI
10.1021/bi500340z
PMC
PMC4071092
OpenAlex
W2029424321
Cited by
30
References
8
Citations by year
2026: 3 · 2025: 4 · 2024: 2 · 2023: 3 · 2020: 5 · 2019: 2 · 2017: 6 · 2016: 2 · 2015: 1 · 2014: 2
FWCI
1.38
Type
journal article
Language
en
Open access
Licence
Locations
  • Biochemistry (publishedVersion, other-oa)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • Japan Society for the Promotion of Science (25293006, 25670014)
  • National Heart, Lung, and Blood Institute (R01HL056083, HL-56083)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
Keywords
apolipoprotein E, APOE ε4, APOE3, domain stability, helix bundle, C-terminal domain, lipid binding, lipoprotein binding, protein unfolding, Alzheimer's disease, cardiovascular risk
MeSH terms
Animals, Apolipoproteins E, Dimyristoylphosphatidylcholine, Humans, Lipids, Lipoproteins, VLDL, Protein Binding, Triolein, Protein Structure, Tertiary, Protein Isoforms, Mice, Apolipoprotein E3, Apolipoprotein E4, Protein Stability

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