Oxidized Proteins Information

Oxidatively Modified Proteins, Lipoproteins and Antibodies

Atherosclerosis is the leading cause of death in the United States, resulting in approximately 500,000 deaths annually. This disease causes significant morbidity, and the associated economic costs have been estimated at $60 billion a year. A large and growing body of evidence implicates the oxidation of low-density lipoproteins (LDLs) in the initiation of atherogenesis. Diabetes mellitus is a chronic disease characterized by abnormal sugar, fat, and protein metabolism. These abnormalities cause the development of atherosclerotic disease, sometimes called hardening of the arteries. Nearly 80% of people with diabetes suffer with heart disease, which is the leading cause of death in diabetics. Low-Density Lipoproteins, LDL or “bad cholesterol” seems to be involved in the development of atherosclerosis. Thus, diabetes is a strong, independent risk factor for atherosclerosis, the major cause of morbidity and mortality in patients with diabetes. To serve our customers to study in the field of oxidation related diseases, we are proud to offer the following oxidatively modified reagent and antibodies.

  1. Cu2+ oxidized LDL

The oxidation of LDL is accelerated significantly by metal ions and is inhibited by chelating agents. Cu2+ oxidized LDL exhibited chemical and biological properties similar (Yang et al., 1999a; Steinbrecher, 1987). LDL oxidized or otherwise modified in vitro, including Cu-Ox-LDL and MM-LDL and lysophosphatidylcholine, a major lipid component of oxidized LDL, increase the expression of cell adhesion molecules on endothelial cells, which support the adhesion of monocytes and lymphocytes (Kume et al., 1992; Khan et al., 1995; Shih et al., 1999). Structure of in vitro oxidized LDL have been widely studied (Valentinova et al., 1994, Burlet et al., 1995, Mark et al., 1996, Yang et al., 1997, Yang et al., 1999b, Yang et al., 2001.).

  1. Anti-Nitrotyrosine antibodies

Peroxynitrite (ONOO-) is a powerful oxidant, and nitrating species are formed by the reaction of nitric oxide with superoxide (Beckman et al., 1992). Peroxynitrite is a relatively selective oxidant and modifies tyrosine either in the free or the protein-bound form to create nitrotyrosines, leaving detectable footprints of oxidation in vivo (Beckman et al., 1994). Antibodies raised against nitrotyrosine can be used to detect the existence of nitrotyrosine in tissues.


  1. Anti-Malondialdehyde (anti-MDA) antibodies

An elevated plasma level of atherogenic MDA-LDL is a marker for unstable atherosclerotic cardiovascular disease. MDA antibody can be used to detect the existence of MDA modified LDL in circulating system (Yang et al. 2003; Berlett and Stadtman 1997).


  1. Anti-Carboxymethyl (anti-CML) antibodies

The initial step in AGE formation is the nonenzymatic attachment of sugar aldehydes or ketones to the side chains of lysine, arginine, and possibly histidine (Vlassara et al., 1994). The lysineε -amino-derived glycation product, or Schiff base, rearranges to form a more stable amino ketone intermediate known as the Amadori product. The presence of the Amadori product (Nagai et al., 1997), indicative of active glycation, can be demonstrated by its reduction to a stable epimeric mixture of 1-glycitol-lysine and 1-mannitol-lysine, known collectively as hexitol-lysine (HL). Immunocytochemical and biochemical studies have suggested that one particular AGE, Nε-(Carboxymethyl)lysine (CML) is the major AGE that accumulates in vivo (Ikeda et al., 1996; Schleicher et al. 1997). Elevated serum levels of CML are detected in patients with diabetes mellitus (Schleicher et al., 1997) and CML is increased in the vascular tissues of diabetic rodents and humans (Kume et al., 1995; Meng et al., 1998).



  1. Anti-4-Hydroxynonenal (anti-4-HNE) antibodies

Modifications on lysine residues, with formation of carboxylmethyl lysine (CML), malondialdehyde (MDA) and hexitol-lysine are advanced glycation end-products (AGE), and the coupling with reactive aldehyde compounds, such as 4-hydroxynonenal (4-HNE) may appear from lipid oxidation (Guichardant et al., 1998). These modifications feature the oxidative byproducts which react with NH2 groups and form Schiff-base adducts (Mark et al., 1996). LDL treated with 4-HNE or oxidatively modified by Cu++ or by cultured endothelial cells give rise to Michael addition-type 4-HNE adducts that are recognized by 4-HNE-specific antibodies.



  1. 6. Anti-Carbamyl (anti-CBL) antibodies

Carbamylation is a spontaneous nonenzymatic modification of proteins and amino acids by urea-derived isocyanate, which is generally present in human serum and is increased in uremic patients (Kraus and Kraus, 2001). Proteins carbamylated at lysine or the terminal protein amino acids may play an important role in Chronic renal failure subjects in the development of atherosclerosis and cardiovascular diseases (Culleton et al., 1999).



Beckman, Joseph S., Harry Ischiropoulos, Ling Zhu, Mark Van Der Woerd, Craig Smith, Jun Chen, Joseph Harrison, James C. Martin, and Michael Tsai. "Kinetics of Superoxide Dismutase- and Iron-catalyzed Nitration of Phenolics by Peroxynitrite." Archives of Biochemistry and Biophysics 298.2 (1992): 438-45.

Beckmann, Joseph S., Yao Zu Ye, Peter G. Anderson, Jun Chen, Mary Ann Accavitti, Margaret M. Tarpey, and C. Roger White. "Extensive Nitration of Protein Tyrosines in Human Atherosclerosis Detected by Immunohistochemistry." Biological Chemistry Hoppe-Seyler 375.2 (1994): 81-88.

Berlett, B. S., and E. R. Stadtman. "Protein Oxidation in Aging, Disease, and Oxidative Stress." Journal of Biological Chemistry 272.33 (1997): 20313-0316.

Burlet, Odile, Chao-Yuh Yang, John R. Guyton, and Simon J. Gaskell. "Tandem Mass Spectrometric Characterization of a Specific Cysteic Acid Residue in Oxidized Human Apoprotein B-100." Journal of the American Society for Mass Spectrometry 6 (1995): 242-47.

Bolgar, M. S., C.-Y. Yang, and S. J. Gaskell. "First Direct Evidence for Lipid/Protein Conjugation in Oxidized Human Low Density Lipoprotein." Journal of Biological Chemistry 271 (1996): 27999-8001.

Culleton, Bruce F., Martin G. Larson, Peter W.f. Wilson, Jane C. Evans, Patrick S. Parfrey, and Daniel Levy. "Cardiovascular Disease and Mortality in a Community-based Cohort with Mild Renal Insufficiency." Kidney International 56 (1999): 2214-219.

Guichardant, Michel, Pascale Taibi-Tronche, Laurent B. Fay, and Michel Lagarde. "Covalent Modifications of Aminophospholipids by 4-hydroxynonenal." Free Radical Biology and Medicine 25.9 (1998): 1049-056.

Ikeda, Kazuyoshi, Takayuki Higashi, Hiroyuki Sano, Yoshiteru Jinnouchi, Masaki Yoshida, Tomohiro Araki, Shoichi Ueda, and Seikoh Horiuchi. "N ε -(Carboxymethyl)lysine Protein Adduct Is a Major Immunological Epitope in Proteins Modified with Advanced Glycation End Products of the Maillard Reaction." Biochemistry 35 (1996): 8075-083.

Khan, B. V., S. S. Parthasarathy, R. W. Alexander, and R. M. Medford. "Modified Low Density Lipoprotein and Its Constituents Augment Cytokine-activated Vascular Cell Adhesion Molecule-1 Gene Expression in Human Vascular Endothelial Cells." Journal of Clinical Investigation 95.3 (1995): 1262-270.

Kume, N., M. I. Cybulsky, and M. A. Gimbrone. "Lysophosphatidylcholine, a Component of Atherogenic Lipoproteins, Induces Mononuclear Leukocyte Adhesion Molecules in Cultured Human and Rabbit Arterial Endothelial Cells." Journal of Clinical Investigation 90.3 (1992): 1138-144.

Kume, S., M. Takeya, T. Mori, N. Araki, H. Suzuki, S. Horiuchi, T. Kodama, Y. Miyauchi, and K. Takahashi. “Immunohistochemical and ultrastructural detection of advanced glycation end products in atherosclerotic lesions of human aorta with a novel specific monoclonal antibody.” Am J Pathol. 147.3 (1995):654-67.

Kraus, Lorraine M., and Alfred P. Kraus. "Carbamoylation of Amino Acids and Proteins in Uremia." Kidney International 78 (2001): 102-07.

Meng, Jing, Noriyuki Sakata, Shigeo Takebayashi, Takashi Asano, Tetsuhiro Futata, Ryoji Nagai, Kazuyoshi Ikeda, Seikoh Horiuchi, Theingi Myint, and Naoyuki Taniguchi. "Glycoxidation in Aortic Collagen from STZ-induced Diabetic Rats and Its Relevance to Vascular Damage." Atherosclerosis 136.2 (1998): 355-65.

Nagai, Ryoji, Kazuyoshi Ikeda, Takayuki Higashi, Hiroyuki Sano, Yoshiteru Jinnouchi, Tomohiro Araki, and Seikoh Horiuchi. "Hydroxyl Radical Mediates Nϵ-(Carboxymethyl)lysine Formation from Amadori Product." Biochemical and Biophysical Research Communications 234.1 (1997): 167-72.

Shih, Peggy T., Mariano J. Elices, Zhuang T. Fang, Tatiana P. Ugarova, Dana Strahl, Mary C. Territo, Joy S. Frank, Nicholas L. Kovach, Carlos Cabanas, Judith A. Berliner, and Devendra K. Vora. "Minimally Modified Low-density Lipoprotein Induces Monocyte Adhesion to Endothelial Connecting Segment-1 by Activating β1 Integrin." Journal of Clinical Investigation 103.5 (1999): 613-25.

Schleicher, E. D., E. Wagner, and A. G. Nerlich. "Increased Accumulation of the Glycoxidation Product N(epsilon)-(carboxymethyl)lysine in Human Tissues in Diabetes and Aging." Journal of Clinical Investigation 99 (1997):457-68.

Steinbrecher, U.P. “Oxidation of human low-density lipoprotein results in derivatization of lysine residues of apolipoprotein B by lipid peroxide decomposition products.” J. Biol. Chem. 262(1987): 3603-3608.

Valentinova, Natalia V., Zi-Wei Gu, Manlan Yang, Elena V. Yanushevskaya, Ilya V. Antonov, John R. Guyton, Charles V. Smith, Jr. Antonio M. Gotto, and Chao-Yuh Yang. "Immunoreactivity of Apolipoprotein B-100 in Oxidatively Modified Low Density Lipoprotein." Biological Chemistry Hoppe-Seyler 375 (1994): 651-58.

Vlassara, H., R. Bucala, and L. Striker. “Pathogenic effects of advanced glycosylation: biochemical, biologic, and clinical implications for diabetes and aging.” Lab Invest. 70.2 (1994):138-51

Yang, Chao-Yuh, Zi-Wei Gu, Manlan Yang, Shen-Nan Lin, Gary Siuzdak, and Charles V. Smith. "Identification of Modified Tryptophan Residues in Apolipoprotein B-100 Derived from Copper Ion-Oxidized Low-Density Lipoprotein." Biochemistry 38 (1999a): 15903-5908.

Yang, Chao-Yuh, Zi-Wei Gu, Manlan Yang, Shen-Nan Lin, Anthony J. Garcia-Prats, Lynette K. Rogers, Stephen E. Welty, and Charles V. Smith. “Selective modification of apoB-100 in the oxidation of low-density lipoprotein by myeloperoxidase in vitro.” Journal of Lipid Research 40 (1999b): 686-98

Yang, Chao-Yuh, Zi-Wei Gu, Hui-Xin Yang, Manlan Yang, Antonio M Gotto, and Charles V Smith. "Oxidative Modifications of APOB-100 by Exposure of Low Density Lipoproteins to HOCl In Vitro." Free Radical Biology and Medicine 23 (1997): 82-89.

Yang, Chao-Yuh, "Isolation, Characterization, and Functional Assessment of Oxidatively Modified Subfractions of Circulating Low-Density Lipoproteins." Arteriosclerosis, Thrombosis, and Vascular Biology 23.6 (2003): 1083-090.

Yang, Chao-Yuh, Jin Wang, Andrew N. Krutchinsky, Brian T. Chait, Joel D. Morrisett, and Charles V. Smith, “Selective oxidation in vitro by myeloperoxidase of the N-terminal amine in apolipoprotein B-100.” Journal of Lipid Research 42 (2001): 1891-896.