ARYA Atherosclerosis Journal

The effect of homocysteine thiolactone on paraoxonase and aryl esterase activity of human serum purified paraoxonase 1 in vitro experiments

Document Type : Original Article(s)

Authors

Elham Moshtaghie 1
Hashem Naieri 1
Ali Asghar Moshtaghie 1
Sedigheh Asgary 2

1 Department of Biochemistry, Islamic Azad University, Falavarjan Branch, Isfahan, Iran

2 Department of Biochemistry, Islamic Azad University, Falavarjan Branch AND Isfahan Cardiovascular Research Center, Cardiovascular Research Institute, Isfahan University of Medical Sciences, Isfahan, Iran

https://doi.org/10.48305/arya.v18i0.2319
Abstract
BACKGROUND: The important role of lipoproteins, particularly low-density lipoprotein (LDL) and high-density lipoprotein (HDL), has been highly regarded among the known causes of cardiovascular disease (CVD). A wide range of risk factors may cause structural and functional changes in lipoprotein particles, resulting in deposition and formation of atherosclerotic plaques. Homocysteine is one of the most important risk factors in heart disease, and its atherosclerotic properties appear to be related to its intermediate metabolite called homocysteine thiolactone (HCTL). The major aim of the present investigation was to study the effect of HCTL in different concentrations (10, 50, and 100 μM) on paraoxonase and aryl esterase activities of purified human serum paraoxonase 1 (PON1) antioxidant enzyme related to HDL, as an extracellular hydrolyzing enzyme of HCTL.
METHODS: In order to purify PON1 enzyme from human serum, three-step chromatographic methods including DEAE Sephadex A50, Sephadex G100, and DEAE Sephadex A50 were used. Protein concentration and paraoxonase and aryl esterase activities of each fraction were measured separately and the highest activities fractions were collected and subsequently pooled together for the next steps. Ultimately, both activities of PON1 in the presence of different concentrations of HCTL were measured in triplicate by spectrophotometry technique.
RESULTS: HCTL at concentrations of 50 and 100 μM decreased both paraoxonase and aryl esterase activities (P < 0.05) in comparison with the control group, which is directly related to the increase in HCTL concentration. However, at a concentration of 10 μM HCTL, no significant difference was observed in both paraoxonase and aryl esterase activities compared to the control group.
CONCLUSION: HCTL is a highly toxic and reactive compound that is produced in all cells. Extracellular enzyme PON1 causes its hydrolysis with high efficiency. The results obtained from the present study showed that paraoxonase and aryl esterase activities decreased in vitro in the presence of HCTL and therefore, HCTL may cause changing in the protein structure of this enzyme. Previous in vivo studies have also shown decrease of PON1 activity in patients with hyperhomocysteinemia.

Keywords

Atherosclerosis
Homocysteine Thiolactone
Paraoxonase 1
  1. Hagstrom E, Roe MT, Hafley G, Neely ML, Sidhu MS, Winters KJ, et al. Association between very low levels of high-density lipoprotein cholesterol and long-term outcomes of patients with acute coronary syndrome treated without revascularization: Insights from the TRILOGY ACS Trial. Clin Cardiol 2016; 39(6): 329-37.
  2. Rizzo M, Otvos J, Nikolic D, Montalto G, Toth PP, Banach M. Subfractions and subpopulations of HDL: An update. Curr Med Chem 2014; 21(25): 2881-91.
  3. Chen NC, Yang F, Capecci LM, Gu Z, Schafer AI, Durante W, et al. Regulation of homocysteine metabolism and methylation in human and mouse tissues. FASEB J 2010; 24(8): 2804-17.
  4. Gurda D, Handschuh L, Kotkowiak W, Jakubowski H. Homocysteine thiolactone and N-homocysteinylated protein induce pro-atherogenic changes in gene expression in human vascular endothelial cells. Amino Acids 2015; 47(7): 1319-39.
  5. Azzini E, Ruggeri S, Polito A. Homocysteine: Its possible emerging role in at-risk population groups. Int J Mol Sci 2020; 21(4): 1421.
  6. Jakubowski H. N-Homocysteinyl-Proteins. In: Jakubowski H, editor. Homocysteine in protein structure/function and human disease: Chemical biology of homocysteine-containing proteins. Vienna, Austria: Springer Vienna; 2013. p. 59-105.
  7. Suszynska-Zajczyk J, Wroblewski J, Utyro O, Luczak M, Marczak L, Jakubowski H. Bleomycin hydrolase and hyperhomocysteinemia modulate the expression of mouse proteins involved in liver homeostasis. Amino Acids 2014; 46(6): 1471-80.
  8. Riise R, Odqvist L, Mattsson J, Monkley S, Abdillahi SM, Tyrchan C, et al. Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes. Sci Rep 2019; 9(1): 20407.
  9. Golmanesh L, Mehrani H, Tabei M. Simple procedures for purification and stabilization of human serum paraoxonase-1. J Biochem Biophys Methods 2008; 70(6): 1037-42.
  10. Yilmaz N. Relationship between paraoxonase and homocysteine: crossroads of oxidative diseases. Arch Med Sci 2012; 8(1): 138-53.
  11. Ivanova EA, Myasoedova VA, Melnichenko AA, Grechko AV, Orekhov AN. Small dense low-density lipoprotein as biomarker for atherosclerotic diseases. Oxid Med Cell Longev 2017; 2017: 1273042.
  12. Liao D, Tan H, Hui R, Li Z, Jiang X, Gaubatz J, et al. Hyperhomocysteinemia decreases circulating high-density lipoprotein by inhibiting apolipoprotein A-I Protein synthesis and enhancing HDL cholesterol clearance. Circ Res 2006; 99(6): 598-606.
  13. Mikael LG, Genest J, Rozen R. Elevated homocysteine reduces apolipoprotein A-I expression in hyperhomocysteinemic mice and in males with coronary artery disease. Circ Res 2006; 98(4): 564-71.
  14. Momin M, Jia J, Fan F, Li J, Dou J, Chen D, et al. Relationship between plasma homocysteine level and lipid profiles in a community-based Chinese population. Lipids Health Dis 2017; 16(1): 54.
  15. Galczynski K, Beltowski J, Nowakowski L, Vasilevska D, Rechberger T, Semczuk A. Serum paraoxonase 1 activity and protein N-homocysteinylation in primary human endometrial cancer. Tumour Biol 2018; 40(9): 1010428318797869.
ARYA Atherosclerosis Journal
Volume 18, Issue 2
February 2022
Pages 1-6

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