Comparison of angiopoietin-like protein 3 and 4 reveals structural and mechanistic similarities.
Angiopoietin-Like Protein 3
Angiopoietin-Like Protein 4
/ chemistry
Angiopoietin-like Proteins
/ chemistry
Coronary Artery Disease
/ blood
Heparin
/ pharmacology
Humans
Lipoprotein Lipase
/ chemistry
Lipoproteins, VLDL
/ chemistry
Protein Binding
/ drug effects
Protein Conformation
Substrate Specificity
Triglycerides
/ blood
enzyme kinetics
heparin-binding protein
lipoprotein lipase (LPL)
lipoprotein metabolism
noncompetitive inhibition
protein purification
small-angle X-ray scattering (SAXS)
very-low-density lipoprotein (VLDL)
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
Historique:
received:
22
09
2020
revised:
07
01
2021
accepted:
15
01
2021
pubmed:
23
1
2021
medline:
27
8
2021
entrez:
22
1
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 33482195
pii: S0021-9258(21)00081-8
doi: 10.1016/j.jbc.2021.100312
pmc: PMC7949051
pii:
doi:
Substances chimiques
ANGPTL3 protein, human
0
Angiopoietin-Like Protein 3
0
Angiopoietin-Like Protein 4
0
Angiopoietin-like Proteins
0
Lipoproteins, VLDL
0
Triglycerides
0
Heparin
9005-49-6
LPL protein, human
EC 3.1.1.34
Lipoprotein Lipase
EC 3.1.1.34
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
100312Subventions
Organisme : NCI NIH HHS
ID : P30 CA016086
Pays : United States
Organisme : NIH HHS
ID : S10 OD018483
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL125654
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL144970
Pays : United States
Organisme : NIGMS NIH HHS
ID : R42 GM128484
Pays : United States
Organisme : NIGMS NIH HHS
ID : P30 GM124169
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM103622
Pays : United States
Organisme : NIH HHS
ID : S10 OD018090
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL094463
Pays : United States
Informations de copyright
Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
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