Comparison of angiopoietin-like protein 3 and 4 reveals structural and mechanistic similarities
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved..
Elevated plasma triglycerides are a risk factor for coronary artery disease, which is the leading cause of death worldwide. Lipoprotein lipase (LPL) reduces triglycerides in the blood by hydrolyzing them from triglyceride-rich lipoproteins to release free fatty acids. LPL activity is regulated in a nutritionally responsive manner by macromolecular inhibitors including angiopoietin-like proteins 3 and 4 (ANGPTL3 and ANGPTL4). However, the mechanism by which ANGPTL3 inhibits LPL is unclear, in part due to challenges in obtaining pure protein for study. We used a new purification protocol for the N-terminal domain of ANGPTL3, removing a DNA contaminant, and found DNA-free ANGPTL3 showed enhanced inhibition of LPL. Structural analysis showed that ANGPTL3 formed elongated, flexible trimers and hexamers that did not interconvert. ANGPTL4 formed only elongated flexible trimers. We compared the inhibition of ANGPTL3 and ANGPTL4 using human very-low-density lipoproteins as a substrate and found both were noncompetitive inhibitors. The inhibition constants for the trimeric ANGPTL3 (7.5 ± 0.7 nM) and ANGPTL4 (3.6 ± 1.0 nM) were only 2-fold different. Heparin has previously been reported to interfere with ANGPTL3 binding to LPL, so we questioned if the negatively charged heparin was acting in a similar fashion to the DNA contaminant. We found that ANGPTL3 inhibition is abolished by binding to low-molecular-weight heparin, whereas ANGPTL4 inhibition is not. Our data show new similarities and differences in how ANGPTL3 and ANGPTL4 regulate LPL and opens new avenues of investigating the effect of heparin on LPL inhibition by ANGPTL3.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:296 |
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| Enthalten in: |
The Journal of biological chemistry - 296(2021) vom: 01. Jan., Seite 100312 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gunn, Kathryn H [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 26.08.2021 Date Revised 04.12.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.jbc.2021.100312 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM320444244 |
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| 100 | 1 | |a Gunn, Kathryn H |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Comparison of angiopoietin-like protein 3 and 4 reveals structural and mechanistic similarities |
| 264 | 1 | |c 2021 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 26.08.2021 | ||
| 500 | |a Date Revised 04.12.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved. | ||
| 520 | |a Elevated plasma triglycerides are a risk factor for coronary artery disease, which is the leading cause of death worldwide. Lipoprotein lipase (LPL) reduces triglycerides in the blood by hydrolyzing them from triglyceride-rich lipoproteins to release free fatty acids. LPL activity is regulated in a nutritionally responsive manner by macromolecular inhibitors including angiopoietin-like proteins 3 and 4 (ANGPTL3 and ANGPTL4). However, the mechanism by which ANGPTL3 inhibits LPL is unclear, in part due to challenges in obtaining pure protein for study. We used a new purification protocol for the N-terminal domain of ANGPTL3, removing a DNA contaminant, and found DNA-free ANGPTL3 showed enhanced inhibition of LPL. Structural analysis showed that ANGPTL3 formed elongated, flexible trimers and hexamers that did not interconvert. ANGPTL4 formed only elongated flexible trimers. We compared the inhibition of ANGPTL3 and ANGPTL4 using human very-low-density lipoproteins as a substrate and found both were noncompetitive inhibitors. The inhibition constants for the trimeric ANGPTL3 (7.5 ± 0.7 nM) and ANGPTL4 (3.6 ± 1.0 nM) were only 2-fold different. Heparin has previously been reported to interfere with ANGPTL3 binding to LPL, so we questioned if the negatively charged heparin was acting in a similar fashion to the DNA contaminant. We found that ANGPTL3 inhibition is abolished by binding to low-molecular-weight heparin, whereas ANGPTL4 inhibition is not. Our data show new similarities and differences in how ANGPTL3 and ANGPTL4 regulate LPL and opens new avenues of investigating the effect of heparin on LPL inhibition by ANGPTL3 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a enzyme kinetics | |
| 650 | 4 | |a heparin-binding protein | |
| 650 | 4 | |a lipoprotein lipase (LPL) | |
| 650 | 4 | |a lipoprotein metabolism | |
| 650 | 4 | |a noncompetitive inhibition | |
| 650 | 4 | |a protein purification | |
| 650 | 4 | |a small-angle X-ray scattering (SAXS) | |
| 650 | 4 | |a very-low-density lipoprotein (VLDL) | |
| 650 | 7 | |a ANGPTL3 protein, human |2 NLM | |
| 650 | 7 | |a Angiopoietin-Like Protein 3 |2 NLM | |
| 650 | 7 | |a Angiopoietin-Like Protein 4 |2 NLM | |
| 650 | 7 | |a Angiopoietin-like Proteins |2 NLM | |
| 650 | 7 | |a Lipoproteins, VLDL |2 NLM | |
| 650 | 7 | |a Triglycerides |2 NLM | |
| 650 | 7 | |a Heparin |2 NLM | |
| 650 | 7 | |a 9005-49-6 |2 NLM | |
| 650 | 7 | |a LPL protein, human |2 NLM | |
| 650 | 7 | |a EC 3.1.1.34 |2 NLM | |
| 650 | 7 | |a Lipoprotein Lipase |2 NLM | |
| 650 | 7 | |a EC 3.1.1.34 |2 NLM | |
| 700 | 1 | |a Gutgsell, Aspen R |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Yongmei |e verfasserin |4 aut | |
| 700 | 1 | |a Johnson, Caitlin V |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Jian |e verfasserin |4 aut | |
| 700 | 1 | |a Neher, Saskia B |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:296 |g year:2021 |g day:01 |g month:01 |g pages:100312 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.jbc.2021.100312 |3 Volltext |
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