Cited by 100Alzheimer's disease research and treatmentsProteoglycans and glycosaminoglycans researchProtein Interaction Studies and Fluorescence Analysis

Interaction of the N-Terminal Domain of Apolipoprotein E4 with Heparin

Dong, Jun, Peters-Libeu, Clare A., Weisgraber, Karl H., Segelke, Brent W., Rupp, Bernhard, Capila, Ishan, Hernáiz, Maria J., LeBrun, Laurie A. +1 more·2001·Biochemistry

Abstract

Apolipoprotein E (apoE) is an important lipid-transport protein in human plasma and brain. It has three common isoforms (apoE2, apoE3, and apoE4). ApoE is a major genetic risk factor in heart disease and in neurodegenerative disease, including Alzheimer's disease. The interaction of apoE with heparan sulfate proteoglycans plays an important role in lipoprotein remnant uptake and likely in atherogenesis and Alzheimer's disease. Here we report our studies of the interaction of the N-terminal domain of apoE4 (residues 1-191), which contains the major heparin-binding site, with an enzymatically prepared heparin oligosaccharide. Identified by its high affinity for the N-terminal domain of apoE4, this oligosaccharide was determined to be an octasaccharide of the structure DeltaUAp2S(1-->[4)-alpha-D-GlcNpS6S(1-->4)-alpha-L-IdoAp2S(1-->](3)4)-alpha-D-GlcNpS6S by nuclear magnetic resonance spectroscopy, capillary electrophoresis, and polyacrylamide gel electrophoresis. Kinetic analysis of the interaction between the N-terminal apoE4 fragment and immobilized heparin by surface plasmon resonance yielded a K(d) of 150 nM. A similar binding constant (K(d) = 140 nM) was observed for the interaction between immobilized N-terminal apoE4 and the octasaccharide. Isothermal titration calorimetry revealed a K(d) of 75 nM for the interaction of the N-terminal apoE fragment and the octasaccharide with a binding stoichiometry of approximately 1:1. Using previous studies and molecular modeling, we propose a binding site for this octasaccharide in a basic residue-rich region of helix 4 of the N-terminal fragment. From the X-ray crystal structure of the N-terminal apoE4, we predicted that binding of the octasaccharide at this site would result in a change in intrinsic fluorescence. This prediction was confirmed experimentally by an observed increase in fluorescence intensity with octasaccharide binding corresponding to a K(d) of approximately 1 microM.

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Crossref · 10.1021/bi002417n

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Authors
Dong, J., Peters-Libeu, C. A., Weisgraber, K. H., Segelke, B. W., Rupp, B., Capila, I., Hernáiz, M. J., LeBrun, L. A., & Linhardt, R. J.
Affiliations
  • University of Iowa (US)
  • Gladstone Institute of Cardiovascular Disease, Cardiovascular Research Institute, and Department of Pathology, University of California, San Francisco, California 94941-9100, Lawrence Livermore National Laboratory, Biology and Biotechnology Research Program, L-452, University of California, Livermore, California 94550, and Departments of Chemistry, Medicinal and Natural Products Chemistry, and Chemical and Biochemical Engineering, University of Iowa, Iowa City, Iowa 52242
Venue
Biochemistry
Published
2001-02-01
Volume
40
Issue
9
Pages
2826-2834
ISSN
  • 0006-2960 (ISSN-L)
  • 1520-4995
OpenAlex
W2111135465
Cited by
99
References
19
Citations by year
2026: 2 · 2025: 4 · 2024: 3 · 2023: 4 · 2022: 4 · 2021: 4 · 2020: 3 · 2019: 4 · 2018: 2 · 2017: 2 · 2016: 4 · 2015: 2 · 2014: 5 · 2013: 2 · 2012: 1
FWCI
3.79
Type
journal article
Language
en
Open access
closed
Locations
  • Biochemistry (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Heart, Lung, and Blood Institute (HL-41633, P01HL041633, HL 52622, R01HL052622)
  • National Institute of General Medical Sciences (GM38060, R01GM038060)
  • National Institute of Neurological Disorders and Stroke (NS35939, R01NS035939)
Topics
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Interaction Studies and Fluorescence Analysis
Keywords
APOE ε4, N-terminal domain, heparin binding, heparan sulfate proteoglycans, binding affinity, binding stoichiometry, intrinsic fluorescence, Alzheimer's disease, atherogenesis
MeSH terms
Animals, Apolipoproteins E, Calorimetry, Carbohydrate Sequence, Heparin, Kinetics, Molecular Sequence Data, Oligosaccharides, Peptide Fragments, Spectrometry, Fluorescence, Swine, Crystallography, X-Ray, Surface Plasmon Resonance, Apolipoprotein E4

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