Acrolein modification impairs key functional features of rat apolipoprotein E: Identification of modified sites by mass spectrometry

Tuyen N. Tran, Malathi G. Kosaraju, Shiori Tamamizu-Kato, Olayemi Akintunde, Ying Zheng, John K. Bielicki, Kent E Pinkerton, Koji Uchida, Yuan Yu Lee, Vasanthy Narayanaswami

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Apolipoprotein E (apoE), an antiatherogenic apolipoprotein, plays a significant role in the metabolism of lipoproteins. It lowers plasma lipid levels by acting as a ligand for the low-density lipoprotein receptor (LDLr) family of proteins, in addition to playing a role in promoting macrophage cholesterol efflux in atherosclerotic lesions. The objective of this study is to examine the effect of acrolein modification on the structure and function of rat apoE and to determine the sites and nature of modification by mass spectrometry. Acrolein is a highly reactive aldehyde, which is generated endogenously as one of the products of lipid peroxidation and is present in the environment in pollutants such as tobacco smoke and heated oils. In initial studies, acrolein-modified apoE was identified by immunoprecipitation using an acrolein-lysine specific antibody in the plasma of 10-week old male rats that were exposed to filtered air (FA) or low doses of environmental tobacco smoke (ETS). While both groups displayed acrolein-modified apoE in the lipoprotein fraction, the ETS group had higher levels in the lipid-free fraction compared with the FA group. This observation provided the rationale to further investigate the effect of acrolein modification on rat apoE at a molecular level. Treatment of recombinant rat apoE with a 10-fold molar excess of acrolein resulted in (i) a significant decrease in lipid-binding and cholesterol efflux abilities, (ii) impairment in the LDLr- and heparin-binding capabilities, and (iii) significant alterations in the overall stability of the protein. The disruption in the functional abilities is attributed directly or indirectly to acrolein modification yielding an aldimine adduct at K149 and K155 (+38); a propanal adduct at K135 and K138 (+56); an Nε-(3- methylpyridinium)lysine (MP-lysine) at K64, K67, and K254 (+76), and an N ε-(3-formyl-3,4-dehydropiperidino)lysine (FDP-lysine) derivative at position K68 (+94), as determined by matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS). The loss of function may also be attributed to alterations in the overall fold of the protein as noted by changes in the guanidine HCl-induced unfolding pattern and to protein cross-linking. Overall, disruption of the structural and functional integrity of apoE by oxidative modification of essential lysine residues by acrolein is expected to affect its role in maintaining plasma cholesterol homeostasis and lead to dysregulation in lipid metabolism.

Original languageEnglish (US)
Pages (from-to)361-375
Number of pages15
JournalBiochemistry
Volume53
Issue number2
DOIs
StatePublished - Jan 21 2014

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Acrolein
Apolipoproteins E
Mass spectrometry
Rats
Mass Spectrometry
Lysine
Tobacco
Smoke
Lipids
LDL Receptors
Cholesterol
Plasmas
Lipoproteins
Proteins
Air
Apolipoproteins
Protein Stability
Macrophages
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Guanidine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tran, T. N., Kosaraju, M. G., Tamamizu-Kato, S., Akintunde, O., Zheng, Y., Bielicki, J. K., ... Narayanaswami, V. (2014). Acrolein modification impairs key functional features of rat apolipoprotein E: Identification of modified sites by mass spectrometry. Biochemistry, 53(2), 361-375. https://doi.org/10.1021/bi401404u

Acrolein modification impairs key functional features of rat apolipoprotein E : Identification of modified sites by mass spectrometry. / Tran, Tuyen N.; Kosaraju, Malathi G.; Tamamizu-Kato, Shiori; Akintunde, Olayemi; Zheng, Ying; Bielicki, John K.; Pinkerton, Kent E; Uchida, Koji; Lee, Yuan Yu; Narayanaswami, Vasanthy.

In: Biochemistry, Vol. 53, No. 2, 21.01.2014, p. 361-375.

Research output: Contribution to journalArticle

Tran, TN, Kosaraju, MG, Tamamizu-Kato, S, Akintunde, O, Zheng, Y, Bielicki, JK, Pinkerton, KE, Uchida, K, Lee, YY & Narayanaswami, V 2014, 'Acrolein modification impairs key functional features of rat apolipoprotein E: Identification of modified sites by mass spectrometry', Biochemistry, vol. 53, no. 2, pp. 361-375. https://doi.org/10.1021/bi401404u
Tran, Tuyen N. ; Kosaraju, Malathi G. ; Tamamizu-Kato, Shiori ; Akintunde, Olayemi ; Zheng, Ying ; Bielicki, John K. ; Pinkerton, Kent E ; Uchida, Koji ; Lee, Yuan Yu ; Narayanaswami, Vasanthy. / Acrolein modification impairs key functional features of rat apolipoprotein E : Identification of modified sites by mass spectrometry. In: Biochemistry. 2014 ; Vol. 53, No. 2. pp. 361-375.
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AU - Akintunde, Olayemi

AU - Zheng, Ying

AU - Bielicki, John K.

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