Cholesterol Crystal Nucleation from Enzymatically Modified Low-Density Lipoproteins: Combined Effect of Sphingomyelinase and Cholesterol Esterase
Abstract
An assay detecting and quantifying cholesterol nucleation from low-density lipoproteins has been established. Förster resonance energy transfer between dehydroergosterol and dansylated lecithin becomes significantly alleviated as a consequence of conucleation of dehydroergosterol and cholesterol. The assay, in combination with dynamic light scattering, absorbance spectroscopy, and fluorescence microscopy, can be used to study aggregation and nucleation in model blood systems. Human plasma LDL was labeled with dehydroergosterol and dansylated lecithin by incubation with donor multilamellar liposomes and isolated by centrifugation. Exposure of labeled LDL (0.5 mg/mL of total lipids) to sphingomyelinase (0.0-0.2 unit/mL) led to modest particle aggregation but produced no changes in energy transfer and no crystallization. However, addition of sphingomyelinase produced significant particle aggregation, nucleation, and crystallization, in a dose-dependent fashion, in samples that were previously treated with the enzyme, cholesterol esterase (0.2 unit/mL). The combination of cholesterol esterase and sphingomyelinase led to a significant alleviation of energy transfer, which preceded by 24 h the appearance of fluorescent, microscopic sterol crystals. These results point to a synergistic effect between cholesterol esterase and sphingomyelinase, suggesting that mere aggregation of LDL is insufficient to promote nucleation, and crystal formation likely proceeds in the intracellular space after LDL uptake by macrophages.
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Modification of plasma lipoproteins in vitro can induce the formation of cholesterol crystals.
“modification of plasma lipoproteins in vitro can induce the formation of cholesterol crystals56,57.”
From Inflammation and its resolution in atherosclerosis: mediators and therapeutic opportunities (2019) · cites this paper as [57]
Cites
Fluorescence Energy Transfer as a Spectroscopic Ruler
Stryer, L. (1978) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.47.070178.004131 · Cited by 2,408 · see citing works
Öörni, K., Pentikäinen, M. O., Ala-Korpela, M., & Kovanen, P. T. (2000) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)31964-7 · Cited by 252 · see citing works
Tulenko, T. N., Chen, M., Mason, P. E., & Mason, R. P. (1998) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)33861-x · Cited by 172 · see citing works
Self-citationFormation of cholesterol monohydrate crystals in macrophage-derived foam cells.
Tangirala, R. K., Jerome, W. G., Jones, N. L., SMALL, D. M., Johnson, W. J., Glick, J. M., Mahlberg, F. H., & Rothblat, G. H. (1994) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)40131-2 · Cited by 147 · see citing works
Reduced uptake of cholesterol esterase-modified low density lipoprotein by macrophages
Aviram, M., Keidar, S., Rosenblat, M., & Brook, G. J. (1991) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)98994-5 · Cited by 30 · see citing works
Ohta, N., & Hatta, I. (2002) · Chemistry and Physics of Lipids · doi:10.1016/s0009-3084(02)00027-0 · Cited by 28 · see citing works
Wrenn, S. P., Kaler, E. W., & Lee, S. P. (1999) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)33391-5 · Cited by 23 · see citing works
Self-citationWrenn, S. P., Gudheti, M. V., Veleva, A. N., Kaler, E. W., & Lee, S. P. (2001) · Journal of Lipid Research · doi:10.1016/s0022-2275(20)31616-3 · Cited by 17 · see citing works
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Inflammation and its resolution in atherosclerosis: mediators and therapeutic opportunities
Bäck, M., Yurdagul, A., Tabas, I., Öörni, K., & Kovanen, P. T. (2019) · Nature Reviews Cardiology · doi:10.1038/s41569-019-0169-2 · Cited by 1,341 · see citing works
Malekmohammad, K., Bezsonov, E. E., & Rafieian‐Kopaei, M. (2021) · Frontiers in Cardiovascular Medicine · doi:10.3389/fcvm.2021.707529 · Cited by 353 · see citing works
Pietsch, M., & Gütschow, M. (2005) · Journal of Medicinal Chemistry · doi:10.1021/jm0508639 · Cited by 106 · see citing works
Acidification of the intimal fluid: the perfect storm for atherogenesis
Öörni, K., Rajamäki, K., Nguyen, S. D., Lähdesmäki, K., Plihtari, R., Lee-Rueckert, M., & Kovanen, P. T. (2015) · Journal of Lipid Research · doi:10.1194/jlr.r050252 · Cited by 101 · see citing works
Snook, C. F., Jones, J., & Hannun, Y. A. (2006) · Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids · doi:10.1016/j.bbalip.2006.06.004 · Cited by 101 · see citing works
Lehti, S., Nguyen, S. D., Belevich, I., Vihinen, H., Heikkilä, H. M., Soliymani, R., Käkelä, R., Saksi, J., Jauhiainen, M., Grabowski, G. A., et al. (2018) · The American Journal of Pathology · doi:10.1016/j.ajpath.2017.09.019 · Cited by 79 · see citing works
In vitro ultrasound-mediated leakage from phospholipid vesicles
Pong, M., Umchid, S., Guarino, A. J., Lewin, P. A., Litniewski, J., Nowicki, A., & Wrenn, S. P. (2006) · Ultrasonics · doi:10.1016/j.ultras.2006.07.021 · Cited by 67 · see citing works
Self-citationNanoscale depth resolution in scanning near-field infrared microscopy
Wollny, G., Bründermann, E., Arsov, Z., Quaroni, L. G., & Havenith, M. (2008) · Optics Express · doi:10.1364/oe.16.007453 · Cited by 38 · see citing works
Lehti, S., Sjövall, P., Käkelä, R., Mäyränpää, M. I., Kovanen, P. T., & Öörni, K. (2015) · American Journal Of Pathology · doi:10.1016/j.ajpath.2015.01.026 · Cited by 31 · see citing works
Liu, F., Wang, Y., & Yu, J. (2023) · Human Immunology · doi:10.1016/j.humimm.2023.06.002 · Cited by 26 · see citing works
Chong, P. L., & Olsher, M. (2004) · Soft Materials · doi:10.1081/smts-200056098 · Cited by 22 · see citing works
Zschörnig, O., Pietsch, M., Süß, R., Schiller, J., & Gütschow, M. (2005) · Journal of Lipid Research · doi:10.1194/jlr.m400265-jlr200 · Cited by 20 · see citing works
Interactions between sphingomyelin and cholesterol in low density lipoproteins and model membranes
Guarino, A. J., Lee, S. P., & Wrenn, S. P. (2005) · Journal of Colloid and Interface Science · doi:10.1016/j.jcis.2005.06.043 · Cited by 17 · see citing works
Self-citationCombination therapy and drug co-delivery systems for atherosclerosis
Dai, Y., Li, Y., Cao, G., Mo, L., Yang, C., Zhu, Y., Guo, Y., Hong, Y., Xu, H., Lu, S., et al. (2025) · Journal of Controlled Release · doi:10.1016/j.jconrel.2025.02.039 · Cited by 19 · see citing works
Imaging and Targeting Coronary Artery Inflammation
West, H. W., & Antoniades, C. A. (2020) · Antioxidants and Redox Signaling · doi:10.1089/ars.2020.8128 · Cited by 10 · see citing works
Guarino, A. J., Tulenko, T. N., & Wrenn, S. P. (2006) · Chemistry and Physics of Lipids · doi:10.1016/j.chemphyslip.2006.02.020 · Cited by 9 · see citing works
Self-citationTroup, G. M., & Wrenn, S. P. (2004) · Chemistry and Physics of Lipids · doi:10.1016/j.chemphyslip.2004.05.001 · Cited by 8 · see citing works
Self-citationω‐Phthalimidoalkyl Aryl Ureas as Potent and Selective Inhibitors of Cholesterol Esterase
Dato, F. M., Sheikh, M., Uhl, R. Z., Schüller, A. W., Steinkrüger, M., Koch, P., Neudörfl, J., Gütschow, M., Goldfuß, B., & Pietsch, M. (2018) · ChemMedChem · doi:10.1002/cmdc.201800388 · Cited by 6 · see citing works
Aggregation kinetics of low density lipoproteins upon exposure to sphingomyelinase
Guarino, A. J., Lee, S. P., Tulenko, T. N., & Wrenn, S. P. (2004) · Journal of Colloid and Interface Science · doi:10.1016/j.jcis.2004.06.066 · Cited by 6 · see citing works
Self-citationPhotophysical and Structural Characterization of Intrinsically Fluorescent Sterol Aggregates
Reinholdt, P., Joensen, L. E., Petersen, D., Szomek, M., Mularski, A., Simonsen, A. C., Kongsted, J., & Wüstner, D. (2021) · The Journal of Physical Chemistry B · doi:10.1021/acs.jpcb.1c00628 · Cited by 5 · see citing works
Lipid Droplet Surface Promotes 3D Morphological Evolution of Non‐Rhomboidal Cholesterol Crystals
Lee, H., Kang, S., & Choi, S. Q. (2024) · Advanced Science · doi:10.1002/advs.202409201 · Cited by 2 · see citing works
Role of CCs and Their Lipoprotein Precursors in NLRP3 and IL-1β Activation
Rajamäki, K., & Öörni, K. (2023) · Contemporary cardiology · doi:10.1007/978-3-031-41192-2_15
Lee, H., Kang, S., & Choi, S. Q. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.07.18.604130
Wu, T., Zhang, L., Tian, J., Lu, Y., Yang, Y. F., Yang, X., Jin, P., Gu, Y., Wang, X., & Lin, Y. (2025) · Lipids in Health and Disease · doi:10.1186/s12944-025-02625-3
Associations of lipids with diseases in large cohort studies and randomized trials
Kaur, R. P., & Palagati, K. (2025) · Elsevier eBooks · doi:10.1016/b978-0-443-19116-9.00040-3
Mandinov, L., & Learn, C. A. (2026) · Reviews in Cardiovascular Medicine · doi:10.31083/rcm52626
- Authors
- Guarino, A. J., Tulenko, T. N., & Wrenn, S. P.
- Affiliations
- Thomas Jefferson University (US)
- Corresponding author
- Wrenn, Steven P.
- ORCID
- Tulenko, Thomas N.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2004-01-22
- Volume
- 43
- Issue
- 6
- Pages
- 1685-1693
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2033565279
- Cited by
- 26
- References
- 8
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 2 · 2023: 2 · 2021: 2 · 2020: 1 · 2019: 1 · 2018: 1 · 2017: 1 · 2015: 1 · 2014: 1
- FWCI
- 2.84
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
- Health Sciences › Medicine › Surgery › Lipoproteins and Cardiovascular Health
- Keywords
- low-density lipoprotein, sphingomyelinase, cholesterol esterase, dehydroergosterol, fluorescence resonance energy transfer, lipoprotein aggregation, macrophage uptake, atherosclerosis, enzyme synergy
- MeSH terms
- Cholesterol, Sterol Esterase, Crystallization, Dansyl Compounds, Drug Synergism, Ergosterol, Humans, Lipoproteins, LDL, Liposomes, Microscopy, Fluorescence, Phosphatidylcholines, Type C Phospholipases, Spectrometry, Fluorescence, Sphingomyelin Phosphodiesterase, Fluorescence Resonance Energy Transfer
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