Redox-proteomics of the effects of homogentisic acid in an in vitro human serum model of alkaptonuric ochronosis

被引:39
作者
Braconi, Daniela [1 ]
Bianchini, Claretta [1 ]
Bernardini, Giulia [1 ]
Laschi, Marcella [1 ]
Millucci, Lia [1 ]
Spreafico, Adriano [2 ]
Santucci, Annalisa [1 ,2 ]
机构
[1] Univ Siena, Dipartimento Biotecnol, I-53100 Siena, SI, Italy
[2] Univ Siena, Ctr Interdipartimentale Studio Biochim Patol Oste, I-53100 Siena, SI, Italy
关键词
LIPID-PEROXIDATION PRODUCTS; APOLIPOPROTEIN-A-I; OXYGEN RADICALS; OXIDATIVE STRESS; PROTEIN DAMAGE; CYSTEINE RESIDUES; N-ACETYLCYSTEINE; PHYTIC ACID; GLUTATHIONE; DEGRADATION;
D O I
10.1007/s10545-011-9377-6
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Alkaptonuria (AKU) is a rare inborn error of metabolism associated with a deficient activity of homogentisate 1,2-dioxygenase (HGO), an enzyme involved in tyrosine and phenylalanine metabolism. Such a deficiency leads to the accumulation of homogentisic acid (HGA) and its oxidized/polymerized products in connective tissues, where melanin-like pigments accumulate (ochronosis). Ochronosis involves especially joints, where an ochronotic arthropathy develops. Little is known on the molecular mechanisms leading to ochronosis and ochronotic arthropathy in AKU. Previous works of ours showed that HGA in vitro propagates oxidative stress through its conversion into benzoquinone acetate (BQA). We hence used an in vitro model consisting of human serum treated with HGA and evaluated the activities of glutathione related anti-oxidant enzymes and levels of compounds indexes of oxidative stress. Proteomics and redox-proteomics were used to identify oxidized proteins and proteins more likely able to bind BQA. Overall, we found that the production of ochronotic pigment in HGA-treated serum is accompanied by lipid peroxidation, decreased activity of the enzyme glutathione peroxidase and massive depletion of thiol groups, together with increased protein carbonylation and thiol oxidation. We also found that BQA was likely to bind carrier proteins and naturally abundant serum proteins, eventually altering their chemico-physical properties. Concluding, our work points towards a critical importance of thiol compounds in counteracting HGA- and BQA- mediated stress in AKU, so that future research for disease biomarkers and pharmacological treatments for AKU and ochronosis will be more easily addressed.
引用
收藏
页码:1163 / 1176
页数:14
相关论文
共 83 条
[1]   Oscillatory oxido-reductive reaction of intracellular hemoglobin in human erythrocyte incubated with o-aminophenol [J].
Akazawa, M ;
Takasaki, M ;
Tomoda, A .
TOHOKU JOURNAL OF EXPERIMENTAL MEDICINE, 2000, 192 (04) :301-312
[2]   Control of extracellular cysteine/cystine redox state by HT-29 cells is independent of cellular glutathione [J].
Anderson, Corinna L. ;
Iyer, Smita S. ;
Ziegler, Thomas R. ;
Jones, Dean P. .
AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY, 2007, 293 (03) :R1069-R1075
[3]  
ANGELES AP, 1989, J RHEUMATOL, V16, P512
[4]   Plasma albumin cysteinylation is regulated by cystathionine β-synthase [J].
Bar-Or, D ;
Curtis, CG ;
Sullivan, A ;
Rael, LT ;
Thomas, GW ;
Craun, M ;
Bar-Oh, R ;
Maclean, KN ;
Kraus, JP .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 325 (04) :1449-1453
[5]  
BELLOMO G, 1990, METHOD ENZYMOL, V186, P627
[6]   Proteomic and Redox-Proteomic Evaluation of Homogentisic Acid and Ascorbic Acid Effects on Human Articular Chondrocytes [J].
Braconi, Daniela ;
Laschi, Marcella ;
Taylor, Adam M. ;
Bernardini, Giulia ;
Spreafico, Adriano ;
Tinti, Laura ;
Gallagher, James A. ;
Santucci, Annalisa .
JOURNAL OF CELLULAR BIOCHEMISTRY, 2010, 111 (04) :922-932
[7]   Evaluation of anti-oxidant treatments in an in vitro model of alkaptonuric ochronosis [J].
Braconi, Daniela ;
Laschi, Marcella ;
Amato, Loredana ;
Bernardini, Giulia ;
Millucci, Lia ;
Marcolongo, Roberto ;
Cavallo, Giovanni ;
Spreafico, Adriano ;
Santucci, Annalisa .
RHEUMATOLOGY, 2010, 49 (10) :1975-1983
[8]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[9]   Cysteine modification by lipid peroxidation products inhibits protein disulfide isomerase [J].
Carbone, DL ;
Doorn, JA ;
Kiebler, Z ;
Petersen, DR .
CHEMICAL RESEARCH IN TOXICOLOGY, 2005, 18 (08) :1324-1331
[10]  
CARLBERG I, 1985, METHOD ENZYMOL, V113, P484