Purification and characterization of Alu I methylase

• Main Authors: Han, Moon H., Yoon, Ho Sup, Suh, Hyang, Yoo, Ook Joon
• Other Authors: School of Biological Sciences
• Format: Article
• Language: English
• Published: 2012
• Subjects: DRNTU::Science::Biological sciences
• Online Access: https://hdl.handle.net/10356/95467; http://hdl.handle.net/10220/8716; http://www.jbmb.or.kr/jbmbonline/18_1/list.html
• Institution: Nanyang Technological University

Alu I methylase has been isolated from 300g (wet weight) cells of Arthrobacter luteus. After ammonium sulfate fractionation, the protein which has methylase activity was purified through phosphocellulose, DEAE-cellulose, Heparin agarose, and Hydroxylapatite column chromatography. The methylated DNA by the purified methylase was resistatnt against Alu I endonuclease. The purified Alu I methylase was essentially homogeneous as judged by 10% SDS-polyacrylamide gel electrophoresis, and the apparent subunit molecular weight was 56,000±1,000. The specific activity of the enzyme was 1.32 × 105 units per mg protein.