Acrolein induces selective protein carbonylation in synaptosomes

Abstract

Acrolein, the most reactive of the α,β-unsaturated aldehydes, is endogenously produced by lipid peroxidation, and has been found increased in the brain of patients with Alzheimer's disease. Although it is known that acrolein increases total protein carbonylation and impairs the function of selected proteins, no study has addressed which proteins are selectively carbonylated by this aldehyde. In this study we investigated the effect of increasing concentrations of acrolein (0, 0.005, 0.05, 0.5, 5, 50 μM) on protein carbonylation in gerbil synaptosomes. In addition, we applied proteomics to identify synaptosomal proteins that were selectively carbonylated by 0.5 μM acrolein. Acrolein increased total protein carbonylation in a dose-dependent manner. Proteomic analysis (2-D electrophoresis followed by mass spectrometry) revealed that tropomyosin-3-gamma isoform 2, tropomyosin-5, β-actin, mitochondrial Tu translation elongation factor (EFTu mt ) and voltagedependent anion channel (VDAC) were significantly carbonylated by acrolein. Consistent with the proteomics studies that have identified specifically oxidized proteins in Alzheimer's Disease (AD) brain, the proteins identified in this study are involved in a wide variety of cellular functions including energy metabolism, neurotransmission, protein synthesis, and cytoskeletal integrity. Our results suggest that acrolein may significantly contribute for oxidative damage in AD brain.Acrolein (2-propen-1-al), the most reactive of the α,β-unsaturated aldehydes (Esterbauer et al., 1991), is a toxic compound found in automobile exhaust gases, overheated cooking oils and a metabolite of cyclophosphamide and allyl alcohol (Halliwell and Gutteridge, 1999). Acrolein can also be endogenously produced as a product of lipid peroxidation (Adams and Klaidman, 1993,Uchida, 1999) and from polyamine metabolism (Takano et al., 2005 Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptNeuroscience. Author manuscript; available in PMC 2008 July 13. Published in final edited form as:Neuroscience. 2007 July 13; 147(3): 674-679. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript . As a consequence, the identification of acrolein adducts has been proposed as a biologic marker for oxidative stress (Kehrer and Biswal, 2000). Accordingly, acrolein levels have been found increased in pathological conditions associated with oxidative stress, such as spinal cord injury (Luo et al., 2005), diabetic nephropathy (Suzuki and Miyata, 1999) and Alzheimer's Disease (AD) (Markesbery and Lovell, 1998,Cal...

Structural modification of soy protein by the lipid peroxidation product acrolein

“…Pocernich, Cardin, Racine, Lauderback, and Butterfield (2001) reported that incubation of synaptosomes (protein concentration of 4 mg/mL) with 50 mmol/L acrolein for 30 min at room temperature led to increase protein carbonyl levels about 4.5-fold compared to the control. Mello et al (2007) found that acrolein induced gerbil synaptosomes (protein concentration of 4 mg/mL) carbonylation linearly with its concentration (0.005-50 mmol/L) at 37 C for 30 min. Acrolein is the most reactive of all a, b-unsaturated aldehydes, which exhibits facile reactivity with the imidazole group of histidine residues, the 3-amino groups of lysine residues, and the sulfhydryl groups of cysteine residues, and eventually incorporates into protein and generates the proteinlinked carbonyl derivative (Esterbauer et al, 1991;LoPachin et al, 2008;Uchida, 1999).…”

Protein modification by acrolein: Relevance to pathological conditions and inhibition by aldehyde sequestering agents

“…ACR selectively modifies synaptosomes proteins related to vital neuronal functions [13]. Proteomic analysis (2-D electrophoresis followed by MS) identified, in synaptosomes isolated from Mongolian gerbils forebrains, tropomyosin-3-g isoform 2, tropomyosin-5, b-actin, mitochondrial Tu translation elongation factor, and voltage-dependent anion channel as the main carbonylated proteins.…”

“…Identifying targets for ACR modification is worthwhile since such knowledge can provide mechanistic insight into cellular or metabolic pathways that are dysregulated by electrophiles, and recent proteomics studies have identified various cytoskeleton-associated proteins as ACR targets [12,13].…”

Aldehydes participation in oxidative stress in rat thymocytes in vitro

“…For example, the most reactive among non-saturated aldehydes ACR is formed endogenously under POL and polyamines metabolism [10]. ACR may induce oxidative stress [11], react with proteins, phospholipids and DNA accompanied by formation of Michael adducts [12].…”

“…ACR may induce oxidative stress [11], react with proteins, phospholipids and DNA accompanied by formation of Michael adducts [12]. Mechanisms of the ACR action are not fully established, but recent researches demonstrate that this carbonyl compound is able to connect and cleavage cell nucleophiles, such as reduced glutathione, lipoic acid and thioredoxin [10]. It can attack free thiol groups of cysteine and γ-amino groups of lysine and histidine with ACR-amino acid adducts and carbonyl groups of proteins [13] formation.…”