title: A glutathione S-transferase (GST) isozyme from broccoli with significant sequence homology to the mammalian theta-class of GSTs.
abstract: A novel glutathione S-transferase (GST) was purified from broccoli (Brassica oleracea var. italica). Partial amino-acid sequencing indicated that the protein shared significant homology with several different plant GSTs from maize, silene, Dianthus, Nicotiana and Triticum, but little homology to yeast (Issatchenkia) GST. One region of the polypeptide near the N-terminal also shared significant homology to a region of rat 5-5, rat 12-12 and human theta-GST (collectively referred to as the theta-GST-class) but little structural homology to the common mammalian cytosolic GSTs (alpha-, mu- or pi-classes). The broccoli GST was retained on a novel membrane based glutathione affinity matrix and displayed activity towards 1-chloro-2,4-dinitro-benzene (CDNB), a general GST substrate, as well as 4-nitrophenethyl bromide, a marker substrate for the theta-class of GSTs. The characteristics of the broccoli GST potentially define it as a member of the theta-class. This is consistent with the view that the theta-class may have arisen prior to the divergence of animals and plants while the mammalian mu-, pi- and alpha-classes evolved after the two kingdoms were established.
authors.affiliation: Analytical Life Sciences Department, Millipore Corporation, Bedford, MA 01727.
authors.forename: M F
authors.initials: MF
authors.lastname: Lopez
authors.forename: W F
authors.initials: WF
authors.lastname: Patton
authors.forename: W B
authors.initials: WB
authors.lastname: Sawlivich
authors.forename: H
authors.initials: H
authors.lastname: Erdjument-Bromage
authors.forename: P
authors.initials: P
authors.lastname: Barry
authors.forename: K
authors.initials: K
authors.lastname: Gmyrek
authors.forename: T
authors.initials: T
authors.lastname: Hines
authors.forename: P
authors.initials: P
authors.lastname: Tempst
authors.forename: W M
authors.initials: WM
authors.lastname: Skea
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001937:Brassica / Q000201:enzymology*; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000145:classification; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000145:classification; D008322:Mammals; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase