Identification and analyses of inhibitors targeting apolipoprotein(a) kringle domains KIV-7, KIV-10, and KV provide insight into kringle domain function
(2020) In The Journal of biological chemistry 295(15). p.5136-5151- Abstract
Increased plasma concentrations of lipoprotein(a) (Lp(a)) are associated with an increased risk for cardiovascular disease. Lp(a) is composed of apolipoprotein(a) (apo(a)) covalently bound to apolipoprotein B of low-density lipoprotein (LDL). Many of apo(a)'s potential pathological properties, such as inhibition of plasmin generation, have been attributed to its main structural domains, the kringles, and have been proposed to be mediated by their lysine-binding sites. However, available small-molecule inhibitors, such as lysine analogs, bind unselectively to kringle domains and are therefore unsuitable for functional characterization of specific kringle domains. Here, we discovered small molecules that specifically bind to the apo(a)... (More)
Increased plasma concentrations of lipoprotein(a) (Lp(a)) are associated with an increased risk for cardiovascular disease. Lp(a) is composed of apolipoprotein(a) (apo(a)) covalently bound to apolipoprotein B of low-density lipoprotein (LDL). Many of apo(a)'s potential pathological properties, such as inhibition of plasmin generation, have been attributed to its main structural domains, the kringles, and have been proposed to be mediated by their lysine-binding sites. However, available small-molecule inhibitors, such as lysine analogs, bind unselectively to kringle domains and are therefore unsuitable for functional characterization of specific kringle domains. Here, we discovered small molecules that specifically bind to the apo(a) kringle domains KIV-7, KIV-10, and KV. Chemical synthesis yielded compound AZ-05, which bound to KIV-10 with a Kd of 0.8 μm and exhibited more than 100-fold selectivity for KIV-10, compared with the other kringle domains tested, including plasminogen kringle 1. To better understand and further improve ligand selectivity, we determined the crystal structures of KIV-7, KIV-10, and KV in complex with small-molecule ligands at 1.6-2.1 Å resolutions. Furthermore, we used these small molecules as chemical probes to characterize the roles of the different apo(a) kringle domains in in vitro assays. These assays revealed the assembly of Lp(a) from apo(a) and LDL, as well as potential pathophysiological mechanisms of Lp(a), including (i) binding to fibrin, (ii) stimulation of smooth-muscle cell proliferation, and (iii) stimulation of LDL uptake into differentiated monocytes. Our results indicate that a small-molecule inhibitor targeting the lysine-binding site of KIV-10 can combat the pathophysiological effects of Lp(a).
(Less)
- author
- publishing date
- 2020-04-10
- type
- Contribution to journal
- publication status
- published
- in
- The Journal of biological chemistry
- volume
- 295
- issue
- 15
- pages
- 5136 - 5151
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- pmid:32132173
- scopus:85083036370
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.RA119.011251
- language
- English
- LU publication?
- no
- additional info
- © 2020 Sandmark et al.
- id
- bb9f995e-06ab-4895-81f3-a2388caabbc9
- date added to LUP
- 2020-07-09 16:28:37
- date last changed
- 2024-05-30 19:04:50
@article{bb9f995e-06ab-4895-81f3-a2388caabbc9,
abstract = {{<p>Increased plasma concentrations of lipoprotein(a) (Lp(a)) are associated with an increased risk for cardiovascular disease. Lp(a) is composed of apolipoprotein(a) (apo(a)) covalently bound to apolipoprotein B of low-density lipoprotein (LDL). Many of apo(a)'s potential pathological properties, such as inhibition of plasmin generation, have been attributed to its main structural domains, the kringles, and have been proposed to be mediated by their lysine-binding sites. However, available small-molecule inhibitors, such as lysine analogs, bind unselectively to kringle domains and are therefore unsuitable for functional characterization of specific kringle domains. Here, we discovered small molecules that specifically bind to the apo(a) kringle domains KIV-7, KIV-10, and KV. Chemical synthesis yielded compound AZ-05, which bound to KIV-10 with a Kd of 0.8 μm and exhibited more than 100-fold selectivity for KIV-10, compared with the other kringle domains tested, including plasminogen kringle 1. To better understand and further improve ligand selectivity, we determined the crystal structures of KIV-7, KIV-10, and KV in complex with small-molecule ligands at 1.6-2.1 Å resolutions. Furthermore, we used these small molecules as chemical probes to characterize the roles of the different apo(a) kringle domains in in vitro assays. These assays revealed the assembly of Lp(a) from apo(a) and LDL, as well as potential pathophysiological mechanisms of Lp(a), including (i) binding to fibrin, (ii) stimulation of smooth-muscle cell proliferation, and (iii) stimulation of LDL uptake into differentiated monocytes. Our results indicate that a small-molecule inhibitor targeting the lysine-binding site of KIV-10 can combat the pathophysiological effects of Lp(a).</p>}},
author = {{Sandmark, Jenny and Tigerström, Anna and Akerud, Tomas and Althage, Magnus and Antonsson, Thomas and Blaho, Stefan and Bodin, Cristian and Boström, Jonas and Chen, Yantao and Dahlén, Anders and Eriksson, Per-Olof and Evertsson, Emma and Fex, Tomas and Fjellström, Ola and Gustafsson, David and Herslöf, Margareta and Hicks, Ryan and Jarkvist, Emelie and Johansson, Carina and Kalies, Inge and Karlsson Svalstedt, Birgitta and Kartberg, Fredrik and Legnehed, Anne and Martinsson, Sofia and Moberg, Andreas and Ridderström, Marianne and Rosengren, Birgitta and Sabirsh, Alan and Thelin, Anders and Vinblad, Johanna and Wellner, Annika U and Xu, Bingze and Östlund-Lindqvist, Ann-Margret and Knecht, Wolfgang}},
issn = {{1083-351X}},
language = {{eng}},
month = {{04}},
number = {{15}},
pages = {{5136--5151}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{The Journal of biological chemistry}},
title = {{Identification and analyses of inhibitors targeting apolipoprotein(a) kringle domains KIV-7, KIV-10, and KV provide insight into kringle domain function}},
url = {{http://dx.doi.org/10.1074/jbc.RA119.011251}},
doi = {{10.1074/jbc.RA119.011251}},
volume = {{295}},
year = {{2020}},
}