Engineering of a metal coordinating site into human glutathione transferase M1-1 based on immobilized metal ion affinity chromatography of homologous rat enzymes
1994 (English)In: Protein Engineering, ISSN 0269-2139, E-ISSN 1460-213X, Vol. 7, no 9, 1115-1119 p.Article in journal (Refereed) Published
Rat glutathione transferase (GST) 3-3 binds to Ni(II)-iminodiacetic acid (IDA)-agarose, whereas other GSTs that are abundant in rat liver do not bind to this immobilized metal ion affinity chromatography (IMAC) adsorbent. Rat GST 3-3 contains two superficially located amino acid residues, His84 and His85, that are suitably positioned for coordination to Ni(II)-IDA-agarose. This particular structural motif is lacking in GSTs that do not bind to the IMAC matrix. Creation of an equivalent His-His structure in the homologous human GST M1-1 by protein engineering afforded a mutant enzyme that displays affinity for Ni(II)-IDA-agarose, in contrast to the wild-type GST M1-1. The results identify a distinct site that is operational in IMAC and suggest an approach to the rational design of novel integral metal coordination sites in proteins.
Place, publisher, year, edition, pages
1994. Vol. 7, no 9, 1115-1119 p.
IdentifiersURN: urn:nbn:se:uu:diva-121128DOI: 10.1093/protein/7.9.1115ISI: A1994PG50700009PubMedID: 7831282OAI: oai:DiVA.org:uu-121128DiVA: diva2:343691