Proteomic Analysis of Human Chylomicron Remnants Isolated by Apolipoprotein B-48 Immunoprecipitation

Overview

Journal J Atheroscler Thromb

Date 2024 Jul 31

PMID 39085140

Authors

Daisaku Masuda

Takeshi Okada

Masami Sairyou

Kazuaki Takafuji

Tohru Ohama

Masahiro Koseki

Makoto Nishida

Yasushi Sakata

Shizuya Yamashita

Affiliations

Soon will be listed here.

Abstract

Aim: Postprandial hypertriglyceridemia (PHTG) is an independent risk factor for coronary heart diseases. PHTG exhibits accumulation of apoB-48 containing chylomicron remnants (CM-Rs) and apoB-100 containing VLDL remnants (VLDL-Rs), which are both known to be atherogenic. However, unlike VLDL-Rs, structural and functional characterization of CM-Rs remains to be elucidated due to challenges in separating CM-Rs from VLDL-Rs. Recently, we successfully isolated CM-Rs and VLDL-Rs utilizing anti-apoB-48 or apoB-100 specific antibodies. This study aimed to characterize the proteome of CM-Rs along with that of VLDL-Rs.

Methods: Eight healthy subjects were enrolled. Venous blood was drawn 3 hours after high-fat-containing meals. We isolated CM-Rs and VLDL-Rs from sera through combination of ultracentrifugation and immunoprecipitation using apoB-48 or apoB-100 specific antibodies, followed by shotgun proteomic analysis.

Results: We identified 42 CM-Rs or VLDL-Rs-associated proteins, including 11 potential newly identified proteins such as platelet basic protein (PPBP) and platelet factor 4, which are chemokines secreted from platelets. ApoA-I, apoA-IV, and clusterin, which are also known as HDL-associated proteins, were significantly more abundant in CM-Rs. Interestingly, apoC-I, which reduces the activity of lipoprotein lipase and eventually inhibits catabolism of remnant proteins, was also more abundant in CM-Rs. Moreover, we identified proteins involved in complement regulation such as complement C3 and vitronectin, and those involved in acute-phase response such as PPBP, serum amyloid A protein 2, and protein S100-A8, in both CM-Rs and VLDL-Rs.

Conclusions: We have firstly characterized the proteome of CM-Rs. These findings may provide an explanation for the atherogenic properties of CM-Rs.

References

Banfi C, Brioschi M, Barcella S, Wait R, Begum S, Galli S . Proteomic analysis of human low-density lipoprotein reveals the presence of prenylcysteine lyase, a hydrogen peroxide-generating enzyme. Proteomics. 2009; 9(5):1344-52. DOI: 10.1002/pmic.200800566. View

Phillips M, Pullinger C, Kroes I, Kroes J, Hardman D, Chen G . A single copy of apolipoprotein B-48 is present on the human chylomicron remnant. J Lipid Res. 1997; 38(6):1170-7. View

Keller A, Nesvizhskii A, Kolker E, Aebersold R . Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem. 2002; 74(20):5383-92. DOI: 10.1021/ac025747h. View

Proctor S, Mamo J . Intimal retention of cholesterol derived from apolipoprotein B100- and apolipoprotein B48-containing lipoproteins in carotid arteries of Watanabe heritable hyperlipidemic rabbits. Arterioscler Thromb Vasc Biol. 2003; 23(9):1595-600. DOI: 10.1161/01.ATV.0000084638.14534.0A. View

Gofman J, Delalla O, Glazier F, Freeman N, Lindgren F, Nichols A . The serum lipoprotein transport system in health, metabolic disorders, atherosclerosis and coronary heart disease. J Clin Lipidol. 2011; 1(2):104-41. DOI: 10.1016/j.jacl.2007.03.001. View

Shah A, Tan L, Long J, Davidson W . Proteomic diversity of high density lipoproteins: our emerging understanding of its importance in lipid transport and beyond. J Lipid Res. 2013; 54(10):2575-85. PMC: 3770071. DOI: 10.1194/jlr.R035725. View

Fredenrich A, Giroux L, Tremblay M, Krimbou L, Davignon J, Cohn J . Plasma lipoprotein distribution of apoC-III in normolipidemic and hypertriglyceridemic subjects: comparison of the apoC-III to apoE ratio in different lipoprotein fractions. J Lipid Res. 1997; 38(7):1421-32. View

Berbee J, Coomans C, Westerterp M, Romijn J, Havekes L, Rensen P . Apolipoprotein CI enhances the biological response to LPS via the CD14/TLR4 pathway by LPS-binding elements in both its N- and C-terminal helix. J Lipid Res. 2010; 51(7):1943-52. PMC: 2882731. DOI: 10.1194/jlr.M006809. View

Mugii S, Hanada H, Takeoka K, Hidaka Y, Masuda D, Ohama T . [Clinical significance of apolipoprotein B-48 (apoB-48) in patients with thyroid disease]. Rinsho Byori. 2009; 57(11):1058-63. View

10.

Masuda T, Tomita M, Ishihama Y . Phase transfer surfactant-aided trypsin digestion for membrane proteome analysis. J Proteome Res. 2008; 7(2):731-40. DOI: 10.1021/pr700658q. View

11.

Masuda D, Nakagawa-Toyama Y, Nakatani K, Inagaki M, Tsubakio-Yamamoto K, Sandoval J . Ezetimibe improves postprandial hyperlipidaemia in patients with type IIb hyperlipidaemia. Eur J Clin Invest. 2009; 39(8):689-98. DOI: 10.1111/j.1365-2362.2009.02163.x. View

12.

Goldberg I, Eckel R, McPherson R . Triglycerides and heart disease: still a hypothesis?. Arterioscler Thromb Vasc Biol. 2011; 31(8):1716-25. PMC: 3141088. DOI: 10.1161/ATVBAHA.111.226100. View

13.

Nesvizhskii A, Keller A, Kolker E, Aebersold R . A statistical model for identifying proteins by tandem mass spectrometry. Anal Chem. 2003; 75(17):4646-58. DOI: 10.1021/ac0341261. View

14.

Hirata A, Okamura T, Sugiyama D, Kuwabara K, Kadota A, Fujiyoshi A . The Relationship between Very High Levels of Serum High-Density Lipoprotein Cholesterol and Cause-Specific Mortality in a 20-Year Follow-Up Study of Japanese General Population. J Atheroscler Thromb. 2016; 23(7):800-9. PMC: 7399265. DOI: 10.5551/jat.33449. View

15.

Fu X, Gharib S, Green P, Aitken M, Frazer D, Park D . Spectral index for assessment of differential protein expression in shotgun proteomics. J Proteome Res. 2008; 7(3):845-54. DOI: 10.1021/pr070271+. View

16.

Dashty M, Motazacker M, Levels J, de Vries M, Mahmoudi M, Peppelenbosch M . Proteome of human plasma very low-density lipoprotein and low-density lipoprotein exhibits a link with coagulation and lipid metabolism. Thromb Haemost. 2014; 111(3):518-30. DOI: 10.1160/TH13-02-0178. View

17.

GOFMAN J, LINDGREN F, Elliott H . Ultracentrifugal studies of lipoproteins of human serum. J Biol Chem. 1949; 179(2):973-9. View

18.

Hoofnagle A, Heinecke J . Lipoproteomics: using mass spectrometry-based proteomics to explore the assembly, structure, and function of lipoproteins. J Lipid Res. 2009; 50(10):1967-75. PMC: 2739765. DOI: 10.1194/jlr.R900015-JLR200. View

19.

Masuda D, Yamashita S . Postprandial Hyperlipidemia and Remnant Lipoproteins. J Atheroscler Thromb. 2016; 24(2):95-109. PMC: 5305681. DOI: 10.5551/jat.RV16003. View

20.

Murad M, Hazem A, Coto-Yglesias F, Dzyubak S, Gupta S, Bancos I . The association of hypertriglyceridemia with cardiovascular events and pancreatitis: a systematic review and meta-analysis. BMC Endocr Disord. 2012; 12:2. PMC: 3342117. DOI: 10.1186/1472-6823-12-2. View