Johansson A S, Ridderström M, Mannervik B
Department of Biochemistry, Uppsala University, Biomedical Center, Uppsala, Sweden.
Mol Pharmacol. 2000 Mar;57(3):619-24. doi: 10.1124/mol.57.3.619.
gamma-L-Glutamyl-S-(benzyl)-L-cysteinyl-R-(-)-phenylglycine (TER 117) has previously been developed for selective inhibition of human glutathione S-transferase P1-1 (GST P1-1) based on the postulated contribution of this isoenzyme to the development of drug resistance in cancer cells. In the present investigation, the inhibitory effect of TER 117 on the human glyoxalase system was studied. Although designed as an inhibitor specific for GST P1-1, TER 117 also competitively inhibits glyoxalase I (K(I) = 0.56 microM). In contrast, no inhibition of glyoxalase II was detected. Reduced glyoxalase activity is expected to raise intracellular levels of toxic 2-oxoaldehydes otherwise eliminated by glyoxalase I. The resulting toxicity would accompany the potentiation of cytostatic drugs, caused by inhibition of the detoxication effected by GST P1-1. TER 117 was designed for efficient inhibition of the most abundant form GST P1-1/Ile105. Therefore, the inhibitory effect of TER 117 on a second allelic variant GST P1-1/Val105 was also studied. TER 117 was shown to competitively inhibit both GST P1-1 variants. The apparent K(I) values at glutathione concentrations relevant to the intracellular milieu were in the micromolar range for both enzyme forms. Extrapolation to free enzyme produced K(I) values of approximately 0.1 microM for both isoenzymes, reflecting the high affinity of GST P1-1 for the inhibitor. Thus, the allelic variation in position 105 of GST P1-1 does not affect the inhibitory potency of TER 117. The inhibitory effects of TER 117 on GST P1-1 and glyoxalase I activities may act in synergy in the cell and improve the effectiveness of chemotherapy.
γ-L-谷氨酰-S-(苄基)-L-半胱氨酰-R-(-)-苯甘氨酸(TER 117)此前已被开发用于选择性抑制人谷胱甘肽S-转移酶P1-1(GST P1-1),基于该同工酶对癌细胞耐药性发展的假定作用。在本研究中,研究了TER 117对人乙二醛酶系统的抑制作用。尽管TER 117被设计为GST P1-1的特异性抑制剂,但它也竞争性抑制乙二醛酶I(K(I)=0.56微摩尔)。相比之下,未检测到对乙二醛酶II的抑制作用。乙二醛酶活性降低预计会提高细胞内有毒的2-氧代醛的水平,否则这些醛会被乙二醛酶I消除。由此产生的毒性将伴随着细胞生长抑制药物的增效作用,这是由GST P1-1解毒作用的抑制引起的。TER 117被设计用于有效抑制最丰富的形式GST P1-1/Ile105。因此,还研究了TER 117对第二种等位基因变体GST P1-1/Val105的抑制作用。结果表明,TER 117对两种GST P1-1变体均有竞争性抑制作用。在与细胞内环境相关的谷胱甘肽浓度下,两种酶形式的表观K(I)值均在微摩尔范围内。外推至游离酶时,两种同工酶的K(I)值约为0.1微摩尔,反映了GST P1-1对该抑制剂的高亲和力。因此,GST P1-1第105位的等位基因变异不影响TER 117的抑制效力。TER 117对GST P1-1和乙二醛酶I活性的抑制作用可能在细胞中协同发挥作用,提高化疗效果。