Protein-protein interaction affinity chromatography of leukotriene C4 synthase
1995 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 6, no 3, 352-356 p.Article in journal (Refereed) Published
A novel affinity chromatography purification for human leukotriene C4 synthase is described. It is based on a specific interaction between leukotriene C4 synthase and microsomal glutathione S-transferase which occurs in the presence of magnesium ion. Microsomal glutathione S-transferase was immobilized on NHS-activated Sepharose 4B and used as an affinity matrix. Microsomes from 12-O-tetradecanoyl phorbol 13-acetate-treated human erythroleukemia cells were solubilized with taurocholic acid and applied on the affinity matrix at 0.1 M Mg2+ concentration. After washing with a buffer containing Mg2+, the enzyme was eluted with a glutathione-containing buffer lacking Mg2+. This facile one-step procedure gave a 166-fold purification of leukotriene C4 synthase with a yield of 44%. Analyses of proteins specifically adsorbed to the affinity matrix revealed components with apparent molecular weights of 18, 37, 48, and 60 kDa.
Place, publisher, year, edition, pages
Elsevier, 1995. Vol. 6, no 3, 352-356 p.
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:liu:diva-99881DOI: 10.1006/prep.1995.1046ISI: A1995RC77000020PubMedID: 7663172OAI: oai:DiVA.org:liu-99881DiVA: diva2:658728