Department of Plant Pathology, University of California, Davis, CA 95616-8680, USA.
J Microbiol. 2013 Feb;51(1):88-99. doi: 10.1007/s12275-013-2434-0. Epub 2013 Mar 2.
Gamma-glutamyltransferase (GGT, EC 2.3.2.2) cleaves the γ-glutamyl linkage in glutathione (GSH). Ascomycetes in either the Saccharomycotina or the Taphrinomycotina have one to three GGTs, whereas members of the Pezizomycotina have two to four GGTs. A Bayesian analysis indicates there are three well-supported main clades of GGTs in the Ascomycota. 1) A Saccharomycotina and a Taphrinomycotina-specific GGT sub-clade form a yeast main clade. This clade has the three relatively well-characterized fungal GGTs: (Saccharomyces cerevisiae CIS2 and Schizosaccharomyces pombe Ggt1 and Ggt2) and most of its members have all 14 of the highly conserved and critical amino acids that are found in GGTs in the other kingdoms. 2) In contrast, a main clade (GGT3) differs in 11 of the 14 highly conserved amino acids that are found in GGTs in the other kingdoms. All of the 44 Pezizomycotina analyzed have either one or two GGT3s. 3) There is a Pezizomycotina-only GGT clade that has two well-supported sub-clades (GGT1 and GGT2); this clade differs in only two of the 14 highly conserved amino acids found in GGTs in the other kingdoms. Because the Pezizomycotina GGTs differ in apparently critical amino acids from the cross-kingdom consensus, a putative GGT from Colletotrichum graminicola, a member of the Pezizomycotina, was cloned and the protein product was expressed as a secreted protein in Pichia pastoris. A GGT enzyme assay of the P. pastoris supernatant showed that the recombinant protein was active, thereby demonstrating that CgGGT1 is a bona fide GGT.
γ-谷氨酰转移酶(GGT,EC 2.3.2.2)切割谷胱甘肽(GSH)中的γ-谷氨酰键。子囊菌门的 Saccharomycotina 或 Taphrinomycotina 有 1 到 3 种 GGT,而 Pezizomycotina 的成员则有 2 到 4 种 GGT。贝叶斯分析表明,子囊菌门中有三个支持良好的 GGT 主要分支。1)Saccharomycotina 和 Taphrinomycotina 特异性 GGT 亚分支形成一个酵母主要分支。这个分支有三个相对特征明显的真菌 GGT:(酿酒酵母 CIS2 和裂殖酵母 Ggt1 和 Ggt2),其大多数成员都有在其他王国的 GGT 中发现的 14 个高度保守和关键的氨基酸。2)相比之下,一个主要分支(GGT3)在其他王国的 GGT 中发现的 14 个高度保守氨基酸中有 11 个不同。分析的 44 个 Pezizomycotina 中的所有成员都有一个或两个 GGT3。3)有一个仅在 Pezizomycotina 中存在的 GGT 分支,它有两个支持良好的亚分支(GGT1 和 GGT2);这个分支在其他王国的 GGT 中发现的 14 个高度保守氨基酸中有两个不同。由于 Pezizomycotina 的 GGT 在明显关键氨基酸上与跨王国共识不同,因此从 Pezizomycotina 的胶孢炭疽菌克隆了一个假定的 GGT,并在毕赤酵母中表达为分泌蛋白。对毕赤酵母上清液的 GGT 酶测定表明,重组蛋白具有活性,从而证明 CgGGT1 是一种真正的 GGT。