Determination of metallothionein in biological fluids using enzyme-linked immunoassay with commercial antibody

• Autorzy: Milnerowicz H. , Bizon A.
• Języki publikacji: EN

Abstrakty

EN

 Metallothionein (MT) is a low molecular weight cysteine-rich protein with a number of roles in the pro/antioxidant balance and homeostasis of essential metals, such as zinc and copper, and in the detoxification of heavy metals, such as cadmium and mercury. Until now, detection of metallothionein in biological fluids remained difficult because of a lack of a broadly reactive commercial test. Meaningful comparison of the values of metallothionein concentrations reported by different authors using their specific isolation procedures and different conditions of enzyme-linked immunoassay is difficult due to the absence of a reference material for metallothionein. Therefore in the present study, we describe a quantitative assay for metallothionein in biological fluids such as plasma and urine performed by a direct enzyme-linked immunoassay using a commercially; available monoclonal mouse anti-metallothionein clone E9 antibody and commercial standards of metallothionein from rabbit liver and a custom preparation of metallothionein from human liver. The sensitivity of the assay for the standard containing two isoforms MT-I and MT-II from human liver was 140 pg/well. The reactivity of the commercial standards and standards containing two isoforms MT-I and MT-II isolated from human liver in our laboratory with a commercial monoclonal mouse anti-metallothionein clone E9 antibody were similar. This suggests that the described ELISA test can be useful for determination of metallothionein concentration in biological fluids. The concentrations of metallothionein in human plasma, erythrocyte lysate and in urine of smoking and non-smoking healthy volunteers are reported. Tobacco smoking increases the extracellular metallothionein concentration (plasma and urine) but does not affect the intracellular concentration (erythrocyte lysate).

Słowa kluczowe

EN

biological fluid; metallothionein; determination; enzyme-linked immunoassay; commercial antibody; human fluid; cysteine-rich protein; identification; quantification; structural characteristics

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2010
• Tom: 57
• Numer: 1
• Opis fizyczny: p.99-104,fig.,ref.

Twórcy

autor

Milnerowicz H.

• Wroclaw University of Medicine, Wroclaw, Poland

autor

Bizon A.

Bibliografia

• Akintola DF, Sampson B, Fleck A (1995) Development of an enzyme-linked immunosorbent assay for human metallothionein-1 in plasma and urine. J Lab Clin Med 126: 119-127. 
• Andrews GK (2000) Regulation of metallothionein gene expression by oxidative stress and metal ions. Biochem Pharmacol 59: 95-105. 
• Aoki Y, Suzuki KT (1991) Detection of metallothionein by western blotting. Methods Enzymol 205: 108-114. 
• Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.  
• Bremner I, Beattie JH (1990) Metallothionein and the trace minerals. Annu Rev Nutr 10: 63-83. 
• Butler AR (1975) The Jaffé reaction. Identification of the coloured species. Clin Chim Acta 59: 227-232. 
• Dabrio M, Rodriguez AR, Bordin G, Bebianno MJ, De Ley M, Sestakova I, Vasak M, Nordberg M (2002) Recent developments in quantification methods for metallothionein. J Inorg Biochem 88: 123-134. 
• Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82: 70-77. 
• Grider A, Kao KJ, Klein PA, Cousins RJ (1989) Enzyme-linked immunosorbent assay for human metallothionein: correlation of induction with infection. J Lab Clin Med 113: 221-228. 
• Hidalgo J, Dingman A, Garvey JS (1989) Preparative isolation of adult human liver metallothionein isoforms. Rev Esp Fisiol 45: 255-264. 
• Hirauchi T, Okabe M, Nagashima T, Niioka T (1999) Metallothionein quantitation in biological materials by enzyme-linked immunosorbent assay (ELISA) using a commercial monoclonal antibody. Trace Elem Electrolytes 16: 177-182.
• Klaassen CD, Liu J, Choudhuri S (1999) Metallothionein: an intracellular protein to protect against cadmium toxicity. Annu Rev Pharmacol Toxicol 39: 267-294. 
• Krizkova S, Adam V, Kizek R (2009) Study of metallothionein oxidation by using of chip CE. Electrophoresis 30: 4029-4033. 
• Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193: 265-275. 
• Lynes MA, Zaffuto K, Unfricht DW, Marusov G, Samson JS, Yin X (2006) The physiological roles of extracellular metallothionein. Exp Biol Med (Maywood) 231: 1548-1554. 
• McCormick CC, Lin LY (1991) Quantification and identification of metallothionein by gel electrophoresis and silver staining. Methods Enzymol 205: 71-78. 
• Milnerowicz H, Chmarek M, Bukowski R, Milnerowicz S (2004) The concentration of metallothionein in fluid collected intrasurgically from pancreatic cysts of patients with diseases of the pancreas. Farmacja - tradycja i nowoczesność (Suppl. 2), SXIV-P018 (in Polish).
• Milnerowicz H, Chmarek M (2005) Influence of smoking on metallothionein level and other proteins binding essential metals in human milk. Acta Paediatr 94: 402-406. 
• Nakajima K, Suzuki K, Otaki N, Kimura M (1991) Epitope mapping of metallothionein antibodies. Methods Enzymol 205: 174-189. 
• Nordberg GF (2009) Historical perspectives on cadmium toxicology. Toxicol Appl Pharmacol 238: 192-200. 
• Nordberg GF, Garvey JS, Chang CC (1982) Metallothionein in plasma and urine of cadmium workers. Environ Res 28: 179-182.  
• Palmiter RD, Findley SD, Whitmore TE, Durnam DM (1992) MT-III, a brain-specific member of the metallothionein gene family. Proc Natl Acad Sci USA 89: 6333-6337. 
• Park JD, Liu Y, Klaassen CD (2001) Protective effect of metallothionein against toxicity of cadmium ant other metals. Toxicology 163: 93-100. 
• Quaife CJ, Findley SD, Erickson JC, Froelick GJ, Kelly EJ, Zambrowicz BP, Palmiter RD (1994) Induction of a new metallothionein isoform (MT-IV) occurs during differentiation of stratified squamous epithelia. Biochemistry 33: 7250-7259. 
• Ronco AM, Arguello G, Suazo M, Llanos MN (2005) Increased levels of metallothionein in placenta of smokers. Toxicology 208: 133-139. 
• Skrzycki M, Majewska M, Podsiad M, Czeczot H (2009) Expression and activity of superoxide dismutase isoenzymes in colorectal cancer. Acta Biochim Pol 56: 663-670. 
• Summer KH, Klein D (1991) Determination of metallothionein in biological materials. Methods Enzymol 205: 57-60. 
• Swierzcek S, Abuknesha RA, Chivers I, Baranovska I, Cunningham P, Price RG (2004) Enzyme-immunoassay for the determination of metallothionein in human urine: application to environmental monitoring. Biomarkers 9: 331-340. 
• Thomas EA, Bailey LB, Kauwell GA, Lee DY, Cousins RJ (1992) Erythrocyte metallothionein response to dietary zinc in humans. J Nutr 122: 2408-2414. 
• Viarengo A, Burlando B, Ceratto N, Panfoli I (2000) Antioxidant role of metallothioneins: a comparative overview. Cell Mol Biol 46: 407-417. 
• Wang R, Sens D, Albrecht A, Garrett S, Somji S, Sens MA, Lu X (2007) Simple method for identification of metallothionein isoforms in cultured human prostate cells by MALDI-TOF/TOF spectrometry. Anal Chem 79: 4433-4441.