Purification and properties of glutathione transferase from Issatchenkia orientalis

Abstract

Glutathione transferase (GST) (EC 2.5.1.18) was purified from a cell extract of Issatchenkia orientalis, and two GST isoenzymes were isolated. They had molecular weights of 37,500 and 40,000 and were designated GST Y-1 and GST Y-2, respectively. GST Y-1 and GST Y-2 gave single bands with molecular weights of 22,000 and 23,500, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. GST Y-1 and GST Y-2 were immunologically distinguished from each other. GST Y-1 showed specific activity 10.4-times and 6.0-times higher when 1-chloro-2,4-dinitrobenzene and o-dinitrobenzene were used as substrates, respectively, than GST Y-2. GST activity was not detected for either isoenzyme when other substrates such as bromosulfophthalein and trans-4-phenyl-3-buten-2-one were used. GST Y-1 and GST Y-2 had Km values of 0.51 and 0.75 mM for glutathione, respectively, and of 0.16 and 4.01 mM for 1-chloro-2,4-dinitrobenzene. GST Y-1 was significantly inhibited by Cibacron blue 3G-A, and GST Y-2 was significantly inhibited by bromosulfophthalein.

Glutathione Transferase in Bacteria: Subunit Composition and Antigenic Characterization

“…The mammalian enzymes are composed either of two identical subunits (homodimers) or combinations of two non-identical subunits (heterodimers) (Awasthi & Singh, 1985 ; Jakoby & Habig, 1980;Ketterer, 1986;Mannervik, 1985;Mannervik & Danielson, 1988). Homodimeric proteins have also been purified from the fungus Mucor jauanicus (Ando et al, 1988) and the yeast Issatchenkia orientalis (Tamaki et al, 1989). The GST of Tetrahymena thermophyla, on the other hand, has been reported to be a monomer (Overbaugh et al, 1988).…”

“…A GST composed of two subunits of M , 22000 has been isolated from the fungus Mucor javanicus (Ando et al., 1988). Two structurally and antigenically distinct GSTs have been recently purified and characterized from the yeast Issatchenkia orientalis (Tamaki et al, 1989). However, very little information is available on the presence, the subunit composition, the antigenic properties and the role of bacterial GSTs.…”

Evolution of Glutathione Metabolism

“…A single GST isoenzyme was detected in Fusarium oxysporum (212). Two immunologically distinct GST homodimers having subunit molecular weights of 22 and 23.5 kDa by SDS-PAGE were purified from the yeast Zssatchenkis orientalis (213). It thus appears that GSTs are produced in fungi and that more than one isoenzyme may occur; however, the generality of such occurrence needs to be more clearly established.…”

Purification and characterization of a glutathione S-transferase from the fungus Cunninghamella elegans

“…Mammalian cytosolic GSTs have been divided into K-, W-, Z-and a-classes on the basis of primary structure, substrate speci¢city and immunological properties [1^3], and further new classes of GSTs such as c [4], U [5] and j [6] are still being discovered. Only a few GSTs have been reported in fungi [7^9], such as Fusarium oxysporum [10], Issatchenkia orientalis [11], Phanerochaete chrysosporium [12], Yarrowia lipolytica [13], and Mucor circinelloides [14]. Investigation of fungal GSTs may help to achieve a better understanding of GST-related proteins [15,16] and GST evolution [9].…”