Affinity chromatography on immobilized S-adenosyl-l-homocysteine : Purification of a furanocoumarin O-methyltransferase from cell cultures of Ruga graveolens L.

✍ Scribed by Satish K. Sharma; Stewart A. Brown

• Publisher: Elsevier Science
• Year: 1978
• Tongue: English
• Weight: 381 KB
• Volume: 157
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(00)92366-5

✦ Synopsis

To a large extent, the present interest in affinity chromatography for enzyme purification originates from the paper of Cuatrecasas et al.'. This study, in which the name affinity chromatography was used for the first time, stimulated an extensive use of this method in the isolation of enzymes, their inhibitors, antibodies, antigens, nucleic acids and a great number of other products, as evidenced by many literature references. The ever increasing number of commercially available insoluble affinants is the best evidence of the rapid development and important role-of this methodz.

Agarose, a polydextran carrier, is the most common support in affinity chromatography to date. Cuatrecasas 3*a has reported the preparation of a great number of agarose derivatives and various groups can therefore be employed for binding. These derivatives are available now as commercial products and include, for example, AH-Sepharose 4B (agarose with covalently bound 1,Qdiaminohexane) which binds compounds through their carboxyl groups via a soluble carbodiimide. In this communication we wish to report an affinity system in which S-adenosyl-L-homocysteine was coupled to AH-Sepharose 4B by the carbodiimide coupling procedure (Fig. I). The potential usefulness of this affinity system is illustrated by results of its application to the purification of an 0-methyltransferase which catalyses the transfer of a methyl group from S-adenosyl-L-methionine to a phenolic furanocoumarjn in Rura grave-oZenss. Mack and SlaytaP have recently used a similar affinity system for the purification of an indole ethylamine N-methyltransferase from Phauaris tuberosa.