Site-specific mutagenesis of human apolipoprotein E. Receptor binding activity of variants with single amino acid substitutions.
Open accessAbstract
Apolipoprotein (apo) E, an important protein involved in cholesterol transport in the plasma, binds with high specificity and high affinity to the apoB, E (low density lipoprotein) receptor. Several lines of evidence have indicated that key basic residues in the vicinity of residues 140-160 of apoE are important in mediating binding to the receptor. Furthermore, apoE variants exhibiting defective receptor binding are associated with the genetic lipid disorder type III hyperlipoproteinemia. To determine whether other basic amino acids in this region of apoE also affect receptor binding activity, site-specific mutagenesis of apoE in a bacterial expression system was undertaken. This system had been used successfully to produce apoE3 that was structurally and functionally equivalent to human plasma apoE3. Variants of apoE in which neutral amino acids were substituted for basic residues at positions 136, 140, 143, and 150 were produced. The variants all displayed defective binding; their activity ranged from 9 to 52% of normal (a range similar to that seen with naturally occurring variants of human apoE). In addition, to determine whether the conformation of this region is important for receptor binding, we designed variants in which proline was substituted for leucine 144 or alanine 152. Both variants were defective, exhibiting 13 and 27% of normal binding, respectively. In contrast, a double mutant in which arginine was substituted for serine 139 and alanine for leucine 149 displayed slightly enhanced receptor binding activity. These studies confirm that the middle of the apoE molecule is important in receptor binding and indicate that only certain amino acid substitutions in this region interfere with receptor binding activity.
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The APOECh mutation impairs APOE binding to GAG and the APOE receptors.
“The APOECh mutation impairs APOE binding to GAG and the APOE receptors2,30.”
From Resilience to autosomal dominant Alzheimer’s disease in a Reelin-COLBOS heterozygous man (2023) · cites this paper as [30]
Compared with ApoE3, ApoE2 and ApoE3ch show 98% and 60% reductions in LDLR binding, respectively.
“Previous reports showed that compared to ApoE3, ApoE2 and ApoE3ch are associate with a 98% and 60% reductions in LDLR binding, respectively11,14.”
From Resistance to autosomal dominant Alzheimer’s disease in an APOE3 Christchurch homozygote: a case report (2019) · cites this paper as [14]
The Christchurch mutation decreases LDLR binding by ~60%.
“Additionally, this mutation was also reported to decrease LDLR binding by ~60%28.”
From Decreased lipidated ApoE‐receptor interactions confer protection against pathogenicity of ApoE and its lipid cargoes in lysosomes (2025) · cites this paper as [28]
The R136S mutation impairs LDLR binding.
“we confirm the R136S mutation impairs LDLR binding28”
From Decreased lipidated ApoE‐receptor interactions confer protection against pathogenicity of ApoE and its lipid cargoes in lysosomes (2025) · cites this paper as [28]
APOE-R136S has severely reduced binding affinity to heparin.
“Notably, the APOE-R136S has severely reduced binding affinity to heparin, as the mutation occurs within the receptor-binding region of APOE36,45,58.”
From The APOE-R136S mutation protects against APOE4-driven tau pathology, neurodegeneration, and neuroinflammation (2023) · cites this paper as [58]
By substituting a positively charged arginine residue for the uncharged serine, the affinity of the receptor-binding domain of APOE-R136S for negatively charged HSPGs is greatly diminished.
“by substituting a positively charged arginine residue for the uncharged serine, the affinity of the receptor-binding domain of APOE-R136S for negatively charged HSPGs is greatly diminished36,45,58”
From The APOE-R136S mutation protects against APOE4-driven tau pathology, neurodegeneration, and neuroinflammation (2023) · cites this paper as [58]
LDL-receptor binding defects for these apoE variants do not correlate well with the severity of dyslipidemia, indicating that the variants may carry additional properties contributing to pathogenic potential.
“However, the LDL-receptor binding defects for these apoE variants do not correlate well with the severity of dyslipidemia, indicating that these variants may carry additional properties that contribute to their pathogenic potential.”
From Biophysical Analysis of Apolipoprotein E3 Variants Linked with Development of Type III Hyperlipoproteinemia (2011) · cites this paper as [28]
Lipid-bound apoE3[R136S] displays a 60% reduction in LDL receptor binding capacity compared to wild-type apoE3.
“In vitro studies showed that lipid-bound apoE3[R136S] displays a reduction of 60% in LDL receptor binding capacity as compared to wild-type apoE3 [28].”
From Biophysical Analysis of Apolipoprotein E3 Variants Linked with Development of Type III Hyperlipoproteinemia (2011) · cites this paper as [28]
All apoE3 variants studied here demonstrate stronger LDL receptor binding compared to wild-type apoE2.
“In contrast, all apoE3 variants studied here demonstrate stronger LDL receptor binding compared to wild-type apoE2 [28], [30], [34], [35].”
From Biophysical Analysis of Apolipoprotein E3 Variants Linked with Development of Type III Hyperlipoproteinemia (2011) · cites this paper as [28]
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- Authors
- Lalazar, A., Weisgraber, K. H., Rall, S. C., Giladi, H., Innerarity, T. L., Levanon, A. Z., Boyles, J. K., Amit, B., Gorecki, M., & Mahley, R. W.
- Affiliations
- University of California, San Francisco (US)
- ORCID
- Mahley, R W
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1988-03
- Volume
- 263
- Issue
- 8
- Pages
- 3542-3545
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1548248031
- Cited by
- 209
- References
- 27
- Citations by year
- 2026: 3 · 2025: 6 · 2024: 5 · 2023: 6 · 2022: 4 · 2021: 3 · 2020: 5 · 2019: 4 · 2018: 1 · 2017: 1 · 2016: 2 · 2015: 2 · 2014: 4 · 2013: 4 · 2012: 3
- FWCI
- 14.74
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
- Health Sciences › Medicine › Surgery › Lipoproteins and Cardiovascular Health
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Lipid metabolism and disorders
- Keywords
- apolipoprotein E, receptor binding, APOE3, LDL receptor, type III hyperlipoproteinemia, basic residues, amino acid substitutions, proline substitution, protein conformation, cholesterol transport, site-directed mutagenesis
- MeSH terms
- Amino Acid Sequence, Amino Acids, Apolipoproteins E, Base Sequence, Escherichia coli, Kinetics, Molecular Sequence Data, Mutation, Receptors, Cell Surface, Recombinant Proteins, Genetic Variation, Low Density Lipoprotein Receptor-Related Protein-1
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