Methyltransferase Colorimetric Assay Kit

Cayman Chemical Item Number 700140

MT Colorimetric Assay Kit

Methyltransferase Colorimetric Assay Kit

Description

Methylation of key biological molecules and proteins plays important roles in numerous biological systems, including signal transduction, biosynthesis, protein repair, gene silencing, and chromatin regulation.¹ The S-adenosylmethionine (SAM) dependent methyltransferases use SAM, the second most commonly used enzymatic cofactor after ATP. SAM, also known as AdoMet, acts as a donor of a methyl group that is required for the modification of proteins and DNA.² Aberrant levels of SAM have been linked to many abnormalities, including Alzheimer’s Disease, depression, Parkinson’s Disease, multiple sclerosis, liver failure, and cancer.^1,2 Cayman’s Methyltransferase Colorimetric Assay Kit is a continuous enzyme-coupled assay that can continuously monitor SAM-dependent methyltransferases.³ The removal of the methyl group from SAM generates S-adenosylhomocysteine (AdoHcy), which is rapidly converted to S-ribosylhomocysteine and adenine by AdoHcy nucleosidase. This rapid conversion prevents the buildup of AdoHcy and its feedback inhibition on the methylation reaction. Finally, the adenine is converted to hypoxanthine, by adenine deaminase, which in turn is converted to urate and hydrogen peroxide (H₂O₂). The rate of production of H₂O₂ is measured with the colorimetric reagent, 3,5-dichloro-2-hydroxybenzenesulfonic acid, by an increase in absorbance at 500-520 nm. The assay is supplied with AdoHcy as a positive control. The assay can be used with any purified SAM-dependent methyltransferase.

1 Loenen, W.A.M. S-Adenosylmethionine: Jack of all trades and master of everything? Biochem Soc Trans 34(2) 330-333 (2006).

2 Chiang, P.K., Gordon, R.K., Tal, J., et al. S-adenosylmethionine and methylation. FASEB J 10 471-480 (1996).

3 Dorgan, K.M., Wooderchak, W.L., Wynn, D.P., et al. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases. Anal Biochem 350 249-255 (2006).

Synonyms

MT Colorimetric Assay Kit

Stability

6 months

Storage

-80°C

Shipping

Dry ice in continental US; may vary elsewhere

Background Reading

Loenen, W.A.M. S-Adenosylmethionine: Jack of all trades and master of everything? Biochem Soc Trans 34(2) 330-333 (2006).

Couture, J., Collazo, E., Hauk, G., et al. Structural basis for the methylation site specificity of SET7/9. Nat Struct Mol Biol 13(2) 140-146 (2006).

Dorgan, K.M., Wooderchak, W.L., Wynn, D.P., et al. An enzyme-coupled continuous spectrophotometric assay for S-adenosylmethionine-dependent methyltransferases. Anal Biochem 350 249-255 (2006).

Chiang, P.K., Gordon, R.K., Tal, J., et al. S-adenosylmethionine and methylation. FASEB J 10 471-480 (1996).

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700140-1ea

96-Well Solid Plate (Colorimetric Assay)